RU2805081C2 - Compositions and corresponding methods for agriculture - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: method for increasing adaptability of a honeybee to organophosphate insecticides includes administering to the honeybee a composition containing an effective amount of organophosphorus insecticides-metabolizing bacteria formulated with a carrier edible to insects. Bacteria that metabolize organophosphate insecticides are selected from the group consisting of Snodgrassella spp., Bartonella spp., Parasaccharibacter spp. and Lactobacillus spp., and are able to colonize the intestines of the bee, thereby improving its adaptability. The invention also relates to a composition for improving adaptability of a honeybee to organophosphate insecticides, comprising an effective amount of organophosphorus insecticide-metabolizing bacteria formulated with an insect-edible carrier.

EFFECT: change in the level, activity or metabolism of one or more microorganisms living in the host organism, such as a honeybee, thereby increasing adaptability of the honeybee to organophosphate insecticides.

10 cl, 12 tbl, 10 ex, 5 dwg

Description

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims benefit from U.S. Provisional Patent Application No. 62/450017, filed January 24, 2017, and U.S. Provisional Patent Application No. 62/583,736, filed November 9, 2017, the contents of which are incorporated herein by reference in their entirety. volume.

BACKGROUND OF THE ART

Some invertebrates, such as nematodes and arthropods (e.g. insects such as European honey bees ( Apis mellifera ) or silkworms ( Bombyx mori )), are used agriculturally in pollination programs and for pest control, and commercially for the production of commercial products such as honey or silk. For the cultivation of beneficial nematodes and arthropods for agricultural or commercial applications, there is a need in the art for methods of promoting the growth and fitness of beneficial invertebrates.

BRIEF DESCRIPTION OF THE INVENTION

Disclosed herein are compositions and methods for modulating the fitness of invertebrates for agricultural or commercial purposes. The composition contains an agent that alters the level, activity or metabolism of one or more microorganisms living in the host, the change resulting in modulation of the fitness of the host.

In one aspect, this document discloses a method of increasing the fitness of a honey bee, the method comprising administering to the honey bee a composition containing an effective amount of organophosphate insecticide metabolizing bacteria formulated with an insect edible carrier.

In some embodiments, administration involves delivering the composition to a honey bee hive or at least one habitat where a honey bee grows, lives, reproduces, or feeds.

In some embodiments, the composition may be liquid, solid, aerosol, paste, gel, or gas.

In some embodiments, the organophosphate insecticide may be fenitrothion.

In certain embodiments, the carrier may be a seed coating agent.

In some embodiments, a honey bee may be present in a honey bee colony.

In another aspect, provided herein is a composition comprising an effective amount of organophosphorus insecticide-metabolizing bacteria formulated with an insect-edible carrier in the form of a liquid, solid, aerosol, paste, gel, or gas.

In some embodiments of the second aspect, organophosphorus insecticide-metabolizing bacteria metabolize fenitrothion.

In some embodiments of the second aspect, the carrier is a seed coating agent.

In some embodiments of the second aspect, the organophosphorus insecticide-metabolizing bacteria are present at a concentration of at least 100,000 cells/mL (e.g., at least about 100,000 cells/mL, at least about 150,000 cells/mL, at least about 200,000 cells/mL). ml, at least about 250,000 cells/ml, at least about 300,000 cells/ml, at least about 350,000 cells/ml, at least about 400,000 cells/ml, at least about 450,000 cells/ml or at least approximately 500,000 cells/ml).

In yet another aspect, the composition contains an agent that alters the level, activity or metabolism of one or more microorganisms living in the body of an insect host, the change resulting in increased fitness of the insect host.

In another case, the composition contains an agent that changes the level, activity or metabolism of one or more microorganisms living in the body of the nematode host, the change resulting in increased fitness of the nematode host.

In some embodiments of any of the above compositions, the one or more microorganisms may be a bacterium or a fungus that resides in the host. In some embodiments, the host bacterium is at least one bacterium selected from the group consisting of Candidatus spp, Buchenera spp, Blattabacterium spp, Baumania spp, Wigglesworthia spp, Wolbachia spp, Rickettsia spp, Orientia spp, Sodalis spp, Burkholderia spp, Cupriavidus spp, Frankia spp, Snirhizobium spp, Streptococcus spp, Wolinella spp, Xylella spp, Erwinia spp, Agrobacterium spp, Bacillus spp, Paenibacillus spp, Streptomyces spp, Micrococcus spp, Corynebacterium spp, Acetobacter spp, Cyanobacteria spp, Salmonella spp, Rhodococcus spp, Pseudomonas spp, Lactobacillus spp, Enterococcus spp, Alcaligenes spp, Klebsiella spp, Paenibacillus spp, Arthrobacter spp, Corynebacterium spp, Brevibacterium spp, Thermus spp, Pseudomonas spp, Clostridium spp and Escherichia spp . In some embodiments, the host fungus is at least one fungus selected from the group consisting of Candida, Metschnikowia, Debaromyces, Starmerella, Pichia, Cryptococcus, Pseudozyma, Symbiotaphrina bucneri, Symbiotaphrina kochii, Scheffersomyces shehatae, Scheffersomyces stipites , Cryptococcus, Trichosporon, Amylostereum areolatum, Epichloe spp, Pichia pinus, Hansenula capsulate, Daldinia decipien, Ceratocytis spp, Ophiostoma spp and Attamyces bromatificus.

In any of the above compositions, the agent, which may also be referred to herein as a modulating agent, can alter the growth, division, viability, metabolism and/or lifespan of a microorganism residing in the host. In any of the above embodiments, the modulating agent may reduce the viability of one or more microorganisms living in the host. In some embodiments, the modulating agent increases the growth rate or viability of one or more microorganisms living in the host.

In any of the above embodiments, the modulating agent is a phage, polypeptide, small molecule, antibiotic, bacterium, or any combination thereof.

In some embodiments, the phage binds to a cell surface protein in a bacterium residing in the host. In some embodiments, the phage is virulent against a bacterium resident in the host. In some embodiments, the phage is at least one phage selected from the group consisting of Myoviridae, Siphoviridae, Podoviridae, Lipothrixviridae, Rudiviridae, Ampullaviridae, Bicaudaviridae, Clavaviridae, Corticoviridae, Cystoviridae, Fuselloviridae, Gluboloviridae, Guttaviridae, Inoviridae, Leviviridae , Microviridae , Plasmaviridae and Tectiviridae.

In some embodiments, the polypeptide is at least one of a bacteriocin, an R-type bacteriocin, a C-rich nodule peptide, an antimicrobial peptide, a lysine, or a bacteriocyte regulatory peptide.

In some embodiments, the small molecule is a metabolite.

In some embodiments, the antibiotic is a broad-spectrum antibiotic.

In some embodiments, the modulating agent is a naturally occurring bacterium. In some embodiments, the bacterium is at least one bacterium selected from the group consisting of Bartonella apis, Parasaccharibacter apium, Frischella perrara, Snodgrassella alvi, Gilliamela apicola, Bifidobacterium spp, and Lactobacillus spp. In some embodiments, the bacterium is at least one bacterium selected from the group consisting of Candidatus spp, Buchenera spp, Blattabacterium spp, Baumania spp, Wigglesworthia spp, Wolbachia spp, Rickettsia spp, Orientia spp, Sodalis spp, Burkholderia spp, Cupriavidus spp, Frankia spp, Snirhizobium spp, Streptococcus spp, Wolinella spp, Xylella spp, Erwinia spp, Agrobacterium spp, Bacillus spp, Paenibacillus spp, Streptomyces spp, Micrococcus spp, Corynebacterium spp, Acetobacter spp, Cyanobacteria spp, Salmonella spp, Rhodococcus spp, Pseudomonas spp, Lactobacillus spp, Enterococcus spp, Alcaligenes spp, Klebsiella spp, Paenibacillus spp, Arthrobacter spp, Corynebacterium spp, Brevibacterium spp, Thermus spp, Pseudomonas spp, Clostridium spp and Escherichia spp . In certain cases, the bacterium is a naturally occurring bacterium that is capable of breaking down pesticides such as organophosphate insecticides (eg, thiophosphate insecticides, eg, fenitrothion).

In any of the above compositions, host fitness can be measured by survival, reproduction or metabolism of the host. In some embodiments, the modulator modulates host fitness by reducing the host's susceptibility to pesticides (eg, susceptibility to a pesticide listed in Table 12). In some embodiments, the pesticide susceptibility is a bactericide or fungicide susceptibility. In some embodiments, the pesticide susceptibility is a susceptibility to insecticides or nematicides.

In any of the above compositions, the composition may contain a variety of different modulating agents. In some embodiments, the composition contains a modulating agent and a pesticide agent (eg, the pesticide shown in Table 12). In some embodiments, the pesticide agent is a bactericidal or fungicidal agent. In some embodiments, the pesticide agent is an insecticidal or nematicidal agent.

In any of the above compositions, the composition may contain a modulating agent and an agent that increases the growth rate of the crop.

In any of the above compositions, the modulating agent may be associated with the second moiety. In some embodiments, the second moiety is a modulating agent.

In any of the above compositions, the modulating agent may be associated with a targeting domain. In some embodiments, the targeting domain targets the modulatory agent to a target site in the host. In some embodiments, the targeting domain targets the modulating agent to one or more microorganisms residing in the host.

In any of the above compositions, the modulatory agent may comprise an inactivating pre- or prosequence, thereby forming a precursor modulatory agent. In some embodiments, the precursor modulatory agent is converted to an active form in the host.

In any of the above compositions, the modulating agent may contain a linker. In some embodiments, the linker is a cleavable linker.

In any of the above compositions, the composition may further comprise a carrier. In some cases, the carrier may be an agriculturally acceptable carrier.

In any of the above compositions, the composition may further comprise a host bait, an adhesive, or a combination thereof. In some embodiments, the host bait is an edible and/or a chemoattractant.

In any of the above compositions, the composition may be contained at a dose effective to modulate host fitness.

In any of the above compositions, the composition may be formulated for delivery to a microorganism residing in the intestine of a host.

In any of the above compositions, the composition may be formulated for delivery to a microorganism residing in a host bacteriocyte and/or the host gut. In some embodiments, the composition may be formulated for delivery to a plant. In some embodiments, the composition may be formulated for use in a host feeder.

In any of the above compositions, the composition may be formulated as a liquid, powder, granules or nanoparticles. In some embodiments, the composition is formulated as a composition selected from the group consisting of a liposome, a polymer, a peptide-secreting bacterium, and a synthetic nanocapsule. In some embodiments, the synthetic nanocapsule delivers the composition to a target site in the host. In some embodiments, the target site is the host's intestine. In some embodiments, the target site is a bacteriocyte in the host.

In a further aspect, hosts that contain any of the above compositions are also provided herein. In some embodiments, the host is an insect. In some embodiments, the insect assists in pest control, pollination, commercial product development, or a combination thereof. In some embodiments, the insect is a species belonging to Coccinellidae, Carabidae, Mantidae, Syrphidae, Lampyridae, Myrmeleontidae, Chrysopidae, Hemerobiidae, Braconidae, Ichneumonidae, or Odonata. In some embodiments, the insect is a species belonging to Andrenidae, Apidae, Colletidae, Halictidae, or Megachilidae . In some embodiments, the insect is a species belonging to Bombycidae or Saturniidae . In certain embodiments, the insect is a honey bee or silkworm.

In some embodiments, the host is a nematode. In some embodiments, the nematode is a species belonging to Heterorhabditis or Steinernema.

In yet another further aspect, a system for modulating host fitness is also provided herein, comprising a modulating agent that specifically targets a microorganism required for host fitness, wherein the system is effective for modulating host fitness, and wherein the host is an insect or nematode . The modulating agent may include any of the compositions described herein. In some embodiments, the modulating agent is formulated as a powder. In some embodiments, the modulating agent is formulated as a solvent. In some embodiments, the modulating agent is formulated as a concentrate. In some embodiments, the modulating agent is formulated as a diluent. In some embodiments, the modulating agent is prepared for delivery by combining any of the previous compositions with a carrier.

In another aspect, methods of modulating the fitness of an insect or nematode using any of the compositions described herein are also provided. In one embodiment, a method of modulating the fitness of an insect host or a nematode host involves delivering a composition of any of the foregoing to a host, wherein the modulating agent specifically affects one or more microorganisms residing in the host and thereby modulates the fitness of the host. In another case, a method of modulating the diversity of microorganisms in the body of an insect host or nematode host involves delivering the composition of any of the previous paragraphs to the host, wherein the modulating agent specifically affects one or more microorganisms living in the host, and thereby modulates the diversity of microorganisms in host body.

In some embodiments of any of the above methods, the modulating agent may alter the levels of one or more microorganisms found in the host. In some embodiments of any of the above methods, the modulating agent may alter the function of one or more microorganisms living in the host. In some embodiments, the one or more microorganisms may be a bacterium and/or a fungus. In some embodiments, one or more microorganisms are required for host fitness. In some embodiments, one or more microorganisms are required for host survival.

In some embodiments of any of the above methods, the delivery step may include providing the modulating agent in a dose and for a time sufficient to affect one or more microorganisms, thereby modulating the diversity of microorganisms in the host. In some embodiments, the delivery step includes topical application of any of the previous compositions to the plant. In some embodiments, the delivery step includes providing the modulating agent through the genetically modified plant. In some embodiments, the delivery step includes providing the modulatory agent to the host in the form of an edible product. In some embodiments, the delivery step includes obtaining a host carrying the modulating agent. In some embodiments, a host carrying a modulatory agent may transfer the modulatory agent to one or more additional hosts.

In some embodiments of any of the above methods, the composition is effective for increasing the health and/or survival of the host. In some embodiments, the composition is effective for increasing host fitness, increasing host longevity, increasing the rate of effective pollination, increasing host product production, increasing host reproductive rate, or a combination thereof. In some embodiments, the composition is effective in reducing the susceptibility of a host to a pesticide agent (eg, the pesticide listed in Table 12). In certain embodiments, the pesticide is a neonicotinoid (eg, imidacloprid). In certain embodiments, the pesticidal agent is an organophosphorus insecticide (eg, a thiophosphate insecticide, eg, fenitrothion). In some embodiments, the composition is effective for increasing host resistance to an allelochemical produced by the plant. In some embodiments, the allelochemical is toxic to the host prior to delivery of the composition. In some embodiments, the allelochemical is caffeine, soy cystatin N, monoterpenes, diterpene acids, or phenolic compounds.

In some embodiments of any of the above methods, the host is an insect. In some embodiments, the insect assists in pest control, pollination, commercial product development, waste decomposition, or a combination thereof. In some embodiments, the insect is a species belonging to Coccinellidae, Carabidae, Mantidae, Syrphidae, Lampyridae, Myrmeleontidae, Chrysopidae, Hemerobiidae, Braconidae, Ichneumonidae, or Odonata. In some embodiments, the insect is a species belonging to Andrenidae, Apidae, Colletidae, Halictidae, or Megachilidae. In some embodiments, the insect is a species belonging to Bombycidae or Saturniidae . In certain embodiments, the insect is a honey bee or silkworm.

In some embodiments, the host is a nematode. In some embodiments, the nematode is a species belonging to Heterorhabditis or Steinernema .

In some embodiments of any of the above methods, the delivery step includes delivering any of the previous compositions to the plant. In some embodiments, the plant is a crop. In some embodiments, the crop is an unharvested crop at the time of delivery. In some embodiments, the crop is a harvested crop at the time of delivery. In some embodiments, the crop includes harvested fruits or vegetables. In some embodiments, the composition is delivered in an amount and for a period of time effective to enhance the growth rate of the crop. In some embodiments, the crop includes corn, soybean, or wheat plants.

In another aspect, this document also provides screening assays to identify a modulatory agent that modulates host fitness. In one case, a screening assay to identify a modulatory agent that modulates host fitness includes the steps of (a) exposing a microorganism that may reside in the host to one or more candidate modulatory agents and (b) identifying a modulatory agent that enhances host fitness.

In some embodiments of the screening assay, the modulating agent is a microorganism that resides in the host. In some embodiments, the microorganism is a bacterium. In some embodiments, the bacterium resident in the host enhances the fitness of the host. In some embodiments, the bacterium breaks down a pesticide (eg, the pesticide shown in Table 12). In certain embodiments, the pesticide is a neonicotinoid (eg, imidacloprid) or an organophosphate insecticide (eg, a thiophosphate insecticide, eg, fenitrothion). In some embodiments, the bacterium secretes an amino acid. In certain embodiments, the amino acid is methionine.

In some embodiments of the screening assay, the modulating agent negatively affects the microorganism that degrades the allelochemical. In some embodiments, the modulatory agent is a phage, antibiotic, or test compound. In some embodiments, the antibiotic is timentin or azithromycin.

In some embodiments of the screening assay, the host may be an invertebrate. In some embodiments, the invertebrate is an insect or nematode. In certain embodiments, the insect is a honey bee. In other specific embodiments, the insect is a silkworm.

In any of the above screening assay embodiments, host fitness can be modulated by altering the host microbiota.

Definitions

As used herein, the term “bacteriocin” refers to a peptide or polypeptide that has antimicrobial properties. Naturally occurring bacteriocins are produced by certain prokaryotes and act against organisms related to the producing strain, but not against the producing strain itself. Bacteriocins provided herein include, without limitation, naturally occurring bacteriocins, such as bacteriocins produced by bacteria, or derivatives thereof, such as engineered bacteriocins, recombinantly expressed bacteriocins, or chemically synthesized bacteriocins. In some cases, the bacteriocin is a functionally active variant of the bacteriocins described herein. In some cases, the bacteriocin variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a specific region or in the entire sequence, the sequence of a bacteriocin described herein, or a naturally occurring bacteriocin.

As used herein, the term "bacteriocyte" refers to a specialized cell found in certain insects that harbor intracellular bacteria with the properties of symbiotic bacteria.

As used herein, the term "effective amount" refers to an amount of a modulatory agent (eg, phage, lysine, bacteriocin, small molecule, or antibiotic) or composition containing the same agent sufficient to produce a specified result, for example, to enhance or promote the fitness of an organism - host (eg insect); to achieve a target level (eg, a predetermined or threshold level) of concentration of the modulating agent in the target host; to achieve a target level (eg, a predetermined or threshold level) of concentration of the modulating agent in the intestine of the target host; to achieve a target level (eg, a predetermined or threshold level) of concentration of the modulating agent in the target host bacteriocyte; to modulate the level or activity of one or more microorganisms (eg, endosymbionts) in the target host.

As used herein, the term "fitness" refers to the ability of a host organism to survive, grow and/or produce viable offspring. The fitness of an organism may be measured by one or more parameters, including but not limited to lifespan, reproductive rate, mobility, body mass, and/or metabolic rate. Fitness may also be measured based on activity (eg pollination) or yield (eg honey or silk).

As used herein, the term "gut" refers to any part of the host's intestine, including the foregut, midgut, or hindgut of the host.

As used herein, the term “host” refers to an organism (eg, an insect) that hosts microorganisms (eg, endogenous microorganisms, endosymbiotic microorganisms (eg, primary or secondary endosymbionts), commensal organisms, and/or pathogenic microorganisms).

As used herein, the term “enhancing host fitness” or “promoting host fitness” refers to any beneficial change in the physiological processes of the host or any activity performed by the host due to administration of a modulatory agent, including without limitation any one or more of the following desired effects : (1) an increase in the host population by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more; (2) increasing the reproduction rate of the host (e.g., insect, e.g., bee or silkworm) by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more; (3) increasing the motility of the host (e.g. insect, e.g. bee or silkworm) by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99 %, 100% or more; (4) an increase in the body weight of the host (e.g., insect, e.g., bee or silkworm) by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more; (5) increasing the metabolic rate or activity of the host (e.g., insect, such as a bee or silkworm) by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95 %, 99%, 100% or more; (6) increasing the pollination rate (e.g., increasing the number of plants pollinated in a specified amount of time) by a host (e.g., an insect such as a bee or silkworm) by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more; (7) increasing the production of waste products in the host (for example, an insect, such as a bee or silkworm) (for example, honey in a honey bee or silk in a silkworm) by approximately 10%, 20%, 30%, 40%, 50%, 60 %, 70%, 80%, 90%, 95%, 99%, 100% or more; (8) increasing the nutrient content of the host (eg insect) (eg protein, fatty acids or amino acids) by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100% or more; or (9) increasing host resistance to a pesticide (e.g., a neonicotinoid (e.g., imidacloprid) or an organophosphate insecticide (e.g., a thiophosphate insecticide, e.g., fenitrothion)) by approximately 10%, 20%, 30%, 40%, 50%, 60 %, 70%, 80%, 90%, 95%, 99%, 100% or more. An increase in host fitness can be determined in comparison to a host to which the modulating agent has not been administered.

The term "insect" includes any organism belonging to the phylum Arthropoda and the class Insecta or the class Arachnida , at any stage of development, i.e., immature or adult insects.

As used herein, the term “lysin,” also known as endolysin, autolysin, murein hydrolase, peptidoglycan hydrolase, or cell wall hydrolase, refers to a hydrolytic enzyme that can lyse a bacterium by cleaving peptidoglycan in the bacterial cell wall. Lysins as provided herein include, without limitation, naturally occurring lysines, such as lysines produced by phages, lysines produced by bacteria, or derivatives thereof, such as engineered lysines, recombinantly expressed lysines, or chemically synthesized lysines. The functionally active bacteriocin variant may be at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a specific region or throughout the sequence of a synthetic, recombinant or naturally occurring bacteriocin, including any described herein.

As used herein, the term "microorganism" refers to bacteria or fungi. Microorganisms may refer to microorganisms resident in the host (eg, endogenous microorganisms, endosymbiotic microorganisms (eg, primary or secondary endosymbionts)), or microorganisms exogenous to the host, including microorganisms that may act as modulators. As used herein, the term “target microorganism” refers to a microorganism that resides in the host and is exposed to the modulating agent, directly or indirectly.

As used herein, the term "agent" or "modulating agent" refers to an agent that is capable of altering the levels and/or functioning of microorganisms residing in a host (eg, insect) and thereby modulate (eg, enhance) the fitness of the organism. host (for example, an insect).

As used herein, the term “pesticide” or “pesticidal agent” refers to a substance that can be used to control agricultural, environmental and household pests such as insects, fungi, bacteria or viruses. The term "pesticide" is understood to include naturally occurring or synthetic insecticides (larvicides or adulticides), insect growth regulators, acaricides (miticides), nematicides, ectoparasiticides, bactericides, fungicides or herbicides (substances that can be used in agriculture to control or modify plant growth). Additional examples of pesticides or pesticide products are given in Table 12. In some cases, the pesticide is an allelochemical. As used herein, the term “allelochemical” or “allelochemical” means a substance produced by an organism that can affect the physiological function (eg, birth, growth, survival or reproduction) of another organism (eg, an insect host or a nematode host).

As used herein, the term “peptide,” “protein,” or “polypeptide” covers any chain of naturally occurring or non-naturally occurring amino acids (both D- and L-amino acids), regardless of length (e.g., at least 2, 3, 4, 5, 6, 7, 10, 12, 14, 16, 18, 20, 25, 30, 40, 50, 100 or more amino acids), the presence or absence of post-translational modifications (eg, glycosylation or phosphorylation) or the presence, for example, of one or more non-aminoacyl groups (e.g., carbohydrate, lipid, etc.) covalently associated with the peptide, and includes, for example, natural proteins, synthetic or recombinant polypeptides and peptides, hybrid molecules, peptoids or peptidomimetics.

As used herein, the "percentage of identity" between two sequences is determined using the BLAST 2.0 algorithm, which is described in Altschul et al. ( J. Mol. Biol. 215:403-410, 1990). BLAST analysis software is publicly available through the National Center for Biotechnology Information.

As used herein, the term “phage” or “bacteriophage” refers to a virus that infects and replicates in bacteria. Bacteriophages replicate in bacteria after introducing their genome into the cytoplasm and do this using a lytic cycle, which leads to lysis of the bacterial cell, or a lysogenic (non-lytic) cycle, in which the bacterial cell remains intact. The phage may be an isolate of a naturally occurring phage or an engineered phage, including vectors or nucleic acids that encode a partial genome of the phage (e.g., including at least all of the essential genes required for the phage's life cycle in a host bacterium) or the complete phage genome.

As used herein, the term “plant” refers to whole plants, plant organs, plant tissues, plant cells, seeds and their progeny. Plant cells include, but are not limited to, cells from seeds, suspension cultures, embryos, meristems, callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen, or microspores. Plant parts include differentiated or undifferentiated tissues, including but not limited to: roots, stems, shoots, leaves, pollen, seeds, tumor tissue, and various forms of cells and cultures (eg, single cells, protoplasts, embryos, or callus tissue). Plant tissue may be located in a plant or in an organ, tissue, or cell culture of a plant. In addition, the plant may be genetically modified to produce heterologous protein or RNA, for example, of any of the modulating agents in the methods and compositions described herein.

The terms "obtained", "produced" or the like are used to indicate that the claim or embodiment relates to the compound, composition, product, etc. as such, i.e., that the compound, composition, product etc. may be prepared or produced by a method that is described for the manufacture of a compound, composition, product, etc., but the compound, composition, product, etc. may also be prepared or produced by methods other than described. The terms “obtained,” “produced,” or the like indicate that the compound, composition, or product is obtained or produced in the specified manner. It should be understood that the terms “obtained,” “produced,” or the like also disclose the terms “obtained,” “produced,” etc. as a preferred embodiment of “obtained,” “produced,” or the like.

Other characteristics and advantages of the present invention will be apparent from the following detailed description and claims.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

The figures are provided to illustrate one or more features, aspects or embodiments of the present invention and are not intended to be limiting.

In fig. Figure 1 shows a graph showing the time it takes for embryos to reach adulthood in Drosophila melanogaster . Drosophila melanogaster embryos were grown on a diet inoculated with Corynebacterium glutamicum (a glutamate-producing strain - C. glutamicum Glu) or on an axenic diet without any bacteria. The percentage of adults emerging from pupae was measured every 12 hours from the time the first adult emerged. Organisms raised on a diet supplemented with bacteria reach adulthood faster than those raised on a diet without added bacteria.

In fig. Figure 2A is a graph showing the effect of male sex on differences in developmental speed in Drosophila melanogaster . The sex of adults was determined, the appearance of which is shown in Fig. 1, and the rate of their appearance was plotted on the graph.

In fig. Figure 2B is a graph showing the effect of female sex on differences in developmental speed in Drosophila melanogaster . The sex of adults was determined, the appearance of which is shown in Fig. 1, and the rate of their appearance was plotted on the graph. The increase in the rate of development in females due to the presence of bacteria in the diet is significantly greater than in similar males. The beneficial effects of the presence of bacteria in the diet of flies are more pronounced in females compared to males.

In fig. 3 is a graph showing that C. glutamicum strains contributed to the production of larval biomass. Larvae raised on diets supplemented with glutamate- or methionine-producing strains of C. glutamicum are larger than larvae raised on sterile or Escherichia coli- supplemented diets. The body areas of the larvae are measured as the number of pixels in the images of the larvae. Medians and 95% confidence intervals are shown as lines on the graph.

In fig. Figure 4 is a panel of graphs showing the results of the Seahorse extracellular flux assay in relation to bacterial respiration. Bacteria were grown to log phase and loaded onto Seahorse XFe96 plates for transient oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) measurements as described in Methods. Treatments were introduced into the wells after approximately 20 minutes, and the bacteria were monitored for changes in growth. Rifampicin=100 µg/ml; chloramphenicol=25 µg/ml; phages (T7 in the case of E. coli and ΦSmVL-C1 in the case of Serratia marcescens ) were lysates diluted 1:2 or 1:100 in SM buffer. The markings on each line are provided solely as indicators of the condition to which each line corresponds and do not indicate data points.

In fig. Figure 5 is a graph showing that phage antagonizing S. marcescens resulted in reduced fly mortality. All flies inoculated with S. marcescens died within a day, while a significant proportion of flies injected with both S. marcescens and phage survived for five days after treatment. Almost all of the control flies, which were not treated in any way, survived to the end of the experiment. The log-rank test was used to compare curves to determine statistical significance, with an asterisk denoting p < 0.0001.

DETAILED DESCRIPTION OF THE INVENTION

Provided herein are methods and compositions for agricultural use, for example, to alter the level, activity, or metabolism of one or more microorganisms residing in a nematode host or arthropod host (e.g., a honey bee or silkworm), wherein the change leads to increased host fitness. The present invention describes a composition that contains a modulating agent (eg, a phage, a peptide, a small molecule, an antibiotic, or combinations thereof) that can alter the microbiota of the host in a manner that is beneficial to the host. The modulator described herein can be used to enhance the fitness of a number of beneficial nematodes or arthropods, such as honey bees and silkworms used in agriculture and commercial activities, by promoting the achievement of favorable levels of microorganisms, microbial activity, microbial metabolism and/or diversity microorganisms.

The methods and compositions described herein are based in part on examples that illustrate how various agents, such as imidacloprid-degrading microorganisms, fenitrothion-degrading microorganisms, and various phages, can be applied to insect hosts such as honeybees or Drosophila , to indirectly improve the health of these hosts by changing the level, activity or metabolism of microorganisms in the bodies of these hosts. Imidacloprid-degrading microorganisms are an illustrative example of neonicotinoid-degrading microorganisms, and more generally illustrate insecticide- or pesticide-degrading microorganisms. Likewise, fenitrothion-degrading microorganisms are an illustrative example of organophosphorus insecticide-degrading microorganisms, and more generally illustrate insecticide- or pesticide-degrading microorganisms. Based on this, the present invention describes a number of different approaches to the use of agents that alter the level, activity or metabolism of one or more microorganisms living in the host, the change resulting in modulation of host fitness.

I. Hosts

i. Hosts

The host for any of the compositions or methods described herein may be any organism belonging to the phylum Nematoda (e.g., nematodes, e.g., beneficial nematodes) or Arthropoda (e.g., insects, e.g., beneficial insects), including any arthropods, described in this document. In some cases, the host is a beneficial insect or nematode (for example, a pollinator, a natural competitor of a pest, or a producer of substances beneficial to humans). As used herein, the term “beneficial insect” or “beneficial nematode” refers to an insect or nematode that provides benefits (eg, economic and/or environmental) to humans, animals, the ecosystem, and/or the environment. For example, the host may be an insect that is involved in the production of a commercial product, including, but not limited to, insects cultured for food (such as honey from honey bees, such as Apis mellifera ), materials (such as silk from Bombyx mori ), and /or substances (for example, varnish from Laccifer lacca or dyes from Dactylopius coccus and Cynipidae ). In addition, the host may include insects or nematodes that are used in agricultural applications, including insects that assist in crop pollination, seed dispersal, or pest control. In addition, in some cases, the host may be an insect suitable for disposal and/or organic processing of waste (for example, earthworms, termites or Diptera larvae).

In some cases, the host produces a usable product (eg, honey, silk, beeswax, or shellac). In some cases, the host is a bee. Exemplary genera of bees include, but are not limited to , Apis, Bombus, Trigona and Osmia . In some cases, the bee is a honey bee (such as an insect belonging to the genus Apis ) . In some cases, the honey bee is the species Apis mellifera (European or Western honey bee), Apis cerana (Asian, eastern or Himalayan honey bee), Apis dorsata ("giant" honey bee), Apis florea ("red dwarf" honey bee) , Apis andreniformis ("black dwarf" honey bee) or Apis nigrocincta . In some cases, the host is a silkworm. The silkworm may be a species from the family Bombycidae or Saturniidae . In some cases the silkworm is Bombyx mori . In some cases, the host is the varnish bug. The lacquer bug may be a species in the family Kerriidae . In some cases, the lac bug is Kerria lacca .

In some cases, the host helps pollinate the plant (for example, bees, beetles, wasps, flies, butterflies or moths). In some examples, the host that helps pollinate the plant is a beetle. In some cases, the beetle is a species from the family Buprestidae, Cantharidae, Cerambycidae, Chrysomelidae, Cleridae, Coccinellidae, Elateridae, Melandryidae, Meloidae, Melyridae, Mordellidae, Nitidulidae, Oedemeridae, Scarabaeidae , or Staphylinidae . In some cases, the host that helps pollinate the plant is a butterfly or moth (such as Lepidoptera ). In some cases, the butterfly or moth is a species from the family Geometridae, Hesperiidae, Lycaenidae, Noctuidae, Nymphalidae, Papilionidae, Pieridae, or Sphingidae . In some cases, the host that helps pollinate the plant is a fly (such as Diptera ). In some cases, the fly belongs to the family Anthomyiidae , Bibionidae, Bombyliidae, Calliphoridae, Cecidomiidae, Ceratopogonidae, Chironomidae, Conopidae, Culicidae, Dolichopodidae, Empididae, Ephydridae, Lonchopteridae, Muscidae, Mycetophilidae, Phoridae, Simuliidae, Stratiomyidae or Syrphidae . In some cases, the host that helps pollinate the plant is an ant (such as Formicidae ), a sawfly (such as Tenthredinidae ) or a wasp (such as Sphecidae or Vespidae ). In some cases, the host that helps pollinate the plant is a bee. In some cases, the bee belongs to the family Andrenidae, Apidae, Colletidae, Halictidae or Megachilidae.

In some cases, the host contributes to pest control. In some cases, the host that contributes to pest control is a predatory nematode. In specific examples, the nematode is a Heterorhabditis or Steinernema species. In some cases, the host that contributes to pest control is an insect. For example, a pest control host may be a species belonging to the family Braconidae (e.g. parasitoid wasps) , Carabidae (e.g. ground beetles) , Chrysopidae (e.g. lacewings) , Coccinellidae (e.g. ladybugs) , Hemerobiidae (e.g. hemerobes) , Ichneumonidae (e.g. ichneumonid parasites) , Lampyridae (e.g. fireflies) , Mantidae (e.g. mantises) , Myrmeleontidae (e.g. antlions) , Odonata (e.g. heteroptera dragonflies and homoptera damselflies), or Syrphidae (e.g. hoverfly ). In other cases, the pest control host is an insect that competes with an insect considered a pest (eg, a crop pest). For example, the Mediterranean fruit fly Ceratitis capitata is a common pest of fruits and vegetables worldwide. One way to control C. capitata is to release sterilized male insects into the environment to compete with wild males for mating with females. In these cases, the host may be a neutered male belonging to a species that is generally considered a pest.

In some cases, the host promotes the decomposition of waste or organic material. In some examples, the host that promotes the decomposition of waste or organic material is a member of the Coleoptera or Diptera . In some cases, the host belonging to Diptera belongs to the family Calliphoridae, Curtonotidae , Drosophilidae, Fanniidae, Heleomyzidae, Milichiidae, Muscidae, Phoridae, Psychodidae, Scatopsidae, Sepsidae, Sphaeroceridae, Stratiomyidae, Syrphidae, Tephritidae or Ulidiidae . In some cases, the host belonging to Coleoptera belongs to the family Carabidae, Hydrophilidae, Phalacaridae , Ptiliidae or Staphylinidae .

In specific cases, the modulating agents disclosed herein can be used to enhance the fitness of honeybee hosts or silkworm hosts.

ii. Host fitness

The methods and compositions provided herein can be used to enhance the fitness of any of the hosts described herein. An increase in fitness may result from any change in the microorganisms living in the host, the changes being a consequence of the administration of a modulating agent and having a beneficial or beneficial effect. impact on the owner.

In some cases, increased host fitness may manifest itself as improvements in host physiological processes (eg, improved health or survival) due to administration of the modulating agent. In some cases, the fitness of an organism may be measured by one or more parameters, including, but not limited to, reproductive rate, lifespan, mobility, fecundity, body mass, metabolic rate, or activity or survival compared to a host that is not treated with the modulating agent. was introduced. For example, the methods or compositions provided herein may be effective for improving the overall health of the host or for improving the overall survival of the host compared to a host to which the modulating agent has not been administered. In some cases, the improvement in host survival is approximately 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100% compared with a reference level (eg, the level found in a host that is not receiving the modulating agent). In some cases, the methods and compositions are effective in increasing the reproductive rate of a host (eg, the rate of reproduction) compared to a host to which the modulating agent has not been administered. In some cases, the methods and compositions are effective in increasing other physiological parameters such as mobility, body weight, lifespan, fertility, or metabolic rate by about 2%, 5%, 10%, 20%, 30%, 40%, 50 %, 60%, 70%, 80%, 90%, 100%, or more than 100% compared to a reference level (eg, the level found in a host that is not receiving the modulatory agent).

In some cases, an increase in host fitness may be manifested in the form of increased production of a product produced by said host compared to a host to which the modulating agent has not been administered. In some cases, the methods or compositions provided herein may be effective in increasing the production of a product produced by a host described herein (e.g., honey, beeswax, beebread, propolis, silk, or varnish) by approximately 2%, 5 %, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100% above a reference level (e.g., the level found in the host , which does not receive a modulating agent).

In some cases, an increase in host fitness may manifest itself as an increase in the frequency or efficiency of desired activities performed by the host (for example, pollination, pest control, seed dispersal, or breakdown of waste or organic material) compared to a host that has not been administered the modulating agent. . In some cases, the methods or compositions provided herein may be effective in increasing the frequency or effectiveness of desired activities performed by the host (eg, pollination, pest control, seed dispersal, or breakdown of waste or organic material) by approximately 2%, 5% , 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100% above a reference level (e.g., the level found in the host, who is not receiving a modulating agent).

In some cases, an increase in host fitness may be manifested by increased production of one or more nutrients in the host (eg, vitamins, carbohydrates, amino acids, or polypeptides) compared to a host that has not been administered the modulating agent. In some cases, the methods or compositions provided herein may be effective in increasing the production of nutrients in a host (e.g., vitamins, carbohydrates, amino acids, or polypeptides) by about 2%, 5%, 10%, 20%, 30% , 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100% compared to a reference level (eg, the level found in a host that is not receiving a modulating agent). In some cases, the methods or compositions provided herein can provide an increase in the nutrient content of a host by increasing the production of nutrients by one or more microorganisms (eg, endosymbionts) in the host.

In some cases, increased host fitness may manifest itself as decreased host susceptibility to a pesticide (e.g., the pesticide listed in Table 12) and/or increased host resistance to the pesticide (e.g., the pesticide listed in Table 12) relative to the organism- host to whom the modulating agent was not administered. In some cases, the methods or compositions provided herein may be effective in reducing host susceptibility to a pesticide agent (e.g., the pesticide listed in Table 12) by about 2%, 5%, 10%, 20%, 30%, 40 %, 50%, 60%, 70%, 80%, 90%, 100%, or greater than 100% compared to a reference level (eg, the level found in a host that is not receiving a modulating agent). In some cases, the host's susceptibility to a pesticide agent is modified by introducing a modulating agent that degrades the pesticide agent (eg, pesticide-degrading bacteria, such as neonicotinoid-degrading bacteria or organophosphorus insecticide-degrading bacteria). The pesticide agent may be any pesticide agent known in the art, including insecticidal agents. In some cases, the pesticide is a neonicotinoid (eg, imidacloprid) or an organophosphorus insecticide (eg, a thiophosphate insecticide, eg, fenitrothion). In some cases, the methods or compositions provided herein may reduce a host's susceptibility to a pesticide agent (e.g., the pesticide listed in Table 12) by increasing the host's ability to metabolize the pesticide agent or break it down into usable substrates.

In some cases, the host's sensitivity to a pesticide is modified by administering a modulating agent that detoxifies the xenobiotic.

In some cases, increased host fitness may be manifested by decreased host susceptibility to the allelochemical and/or increased host resistance to the allelochemical compared to a host to which the modulating agent was not administered. In some cases, the methods and compositions provided herein may be effective in increasing host resistance to an allelochemical by approximately 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70% , 80%, 90%, 100%, or greater than 100% compared to a reference level (eg, the level found in a host that is not receiving the modulating agent). In some cases, the allelochemical is caffeine, soy cystatin N, monoterpenes, diterpene acids, or phenolic compounds. In some cases, the methods or compositions provided herein can reduce a host's susceptibility to an allelochemical by increasing the host's ability to metabolize the allelochemical or break it down into usable substrates.

In some cases, the methods or compositions provided herein may be effective in increasing host resistance to parasites or pathogens (eg, fungal, bacterial or viral pathogens or parasitic microscopic mites (eg, Varroa destructor mite in honey bees)) by compared with a host organism to which the modulating agent was not administered. In some cases, the methods or compositions provided herein may be effective in increasing host resistance to pathogens or parasites (e.g., fungal, bacterial or viral pathogens or parasitic microscopic mites (e.g., the Varroa destructor mite in honey bees)) to approximately 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or more than 100% above the reference level (e.g. level found in a host that does not receive the modulating agent).

In some cases, increased host fitness may manifest itself in the form of other fitness benefits, such as improved tolerance of certain environmental factors (e.g., tolerance of high or low temperature), improved ability to survive in certain habitats, or improved ability to maintain a particular diet (e.g. improved ability to metabolize soybean compared to corn) compared to a host that was not administered the modulating agent. In some cases, the methods or compositions provided herein may be effective in increasing host fitness in a variety of ways described herein. Additionally, the modulating agent may enhance host fitness in any number of host classes, orders, families, genera, or species (e.g., 1 host species, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 200, 250, 500 or more host species). In some cases, the modulating agent acts on one class, order, family, genus or species of hosts.

Host fitness can be assessed using any standard methods in the art. In some cases, host fitness can be assessed by assessing an individual host. Alternatively, host fitness can be assessed by estimating the host population. For example, an increase in host fitness may manifest itself as an increase in the rate of successful competition relative to other insects, thereby resulting in an increase in host population size.

iii. Invertebrate hosts in agriculture

By enhancing the fitness of beneficial nematodes or insects, the modulating agents provided herein may be effective in promoting the growth of plants that benefit from said hosts. The modulator may be delivered using any of the formulations and delivery methods described herein in an amount and for a period of time effective to enhance the fitness of the hosts of interest, thereby producing a beneficial effect on the plant (e.g., by increasing the rate of crop growth, increasing crop yield, reducing pest infestation and/or reducing damage to plants). This may or may not involve direct application of the modulating agent to the plant. For example, in cases where the original host habitat is different from the region in which the plant grows, the modulating agent may be applied to one of the original host habitat or the plants of interest, or a combination of both.

In some cases, the plant may be an agricultural crop such as a cereal, grain, legume, fruit or vegetable. The compositions described herein can be delivered to a crop at any time before or after harvest of a cereal, grain, legume, fruit or vegetable crop. Crop yield is an indicator often used for example in cereals, grains or legumes, and is usually measured in metric tons per hectare (or kilograms per hectare). Crop yield can also refer to the actual production of seeds by the plant. In some cases, the modulator may be effective in increasing crop yield (e.g., increasing the number of metric tons of cereal, grain, legume, fruit, or vegetable crops harvested per hectare and/or increasing seed production) by approximately 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or more compared to the reference level (for example, in a crop to which the modulating agent was not introduced ).

In some cases, a plant (eg a crop) may be at risk of developing a pest (eg insect) infestation, or may already have developed a pest infestation. The methods and compositions described herein can be used to reduce or prevent pest infestation of such crops by promoting the fitness of beneficial insects that prey on the pests. In some cases, a modulator may be effective in reducing crop infestation (e.g., reducing the number of plants infested, reducing pest population size, or reducing damage to plants) by approximately 2%, 5%, 10%, 20%, 30%, 40 %, 50%, 60%, 70%, 80%, 90%, 100% or more compared to the reference level (eg, in a crop to which the modulating agent was not introduced). In other cases, the modulator may be effective in preventing or reducing the likelihood of crop infestation by approximately 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% , 100% or more compared to the reference level (for example, in a crop to which the modulating agent was not introduced).

The compositions and methods described herein can produce beneficial effects on any suitable plant tissue, including, without limitation, somatic embryos, pollen, leaves, stems, calli, stolons, microtubers and shoots. The methods described herein may include the treatment of angiosperms and gymnosperms such as acacia, alfalfa, apple, apricot, artichoke, ash, asparagus, avocado, banana, barley, bean varieties, beets, birch, beech, blackberries, blueberries, broccoli, varieties of Brussels sprouts, cabbage, canola, cantaloupe, carrots, cassava, cauliflower, cedar, cereal plant, celery, chestnut, cherry, Chinese cabbage, citrus plant, clementine, clover, coffee tree, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, chicory, eucalyptus, fava bean varieties, fennel, fig varieties, fir, geranium, grapes, grapefruit, groundnut varieties, physalis, hemlock, hickory, kale, actinidia, kohlrabi , larch, lettuce, leek, lemon, lime, robinia, pine, curly fern, maize, mango, maple, melon, millet, champignon, mustard, nut-bearing plants, oak, varieties of oats, abelmos edible, onion, orange , ornamental plant or flowering plant or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat-forming plant, pepper, persimmon, pigeon pea, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, Italian chicory, radish, rapeseed, raspberries, rice, rye, sorghum, willow, soybeans, spinach, spruce, zucchini, strawberries, sugar beets, sugar cane, sunflower, sweet potatoes, sweet corn, tangerine, tea bush, tobacco, tomato , woody plants, triticale, turf-forming herbaceous plants, varieties of turnips, cultivated grapes, walnuts, watercress, watermelon, wheat, varieties of yams, yew and zucchini.

II. Target microorganisms

Microorganisms targeted by the modulating agent described herein may include any microorganism found in or on the host, including, without limitation, any bacteria and/or fungi described herein. Microorganisms resident in the host may include, for example, symbiotic microorganisms (eg, endosymbiotic microorganisms that provide beneficial nutrients or enzymes to the host), commensal microorganisms, pathogenic or parasitic microorganisms. A symbiotic microorganism (eg, a bacterium or fungus) may be an obligate symbiont of the host or a facultative symbiont of the host. Microorganisms living in the host body can be acquired through any route of transmission, including vertical, horizontal, or multiple origin transmission.

i. Bacteria

Exemplary bacteria that may be targeted in accordance with the methods and compositions provided herein include, but are not limited to, Xenorhabdus spp, Photorhabdus spp, Candidatus spp, Buchnera spp, Blattabacterium spp, Baumania spp, Wigglesworthia spp, Wolbachia spp, Rickettsia spp, Orientia spp, Sodalis spp, Burkholderia spp, Cupriavidus spp, Frankia spp, Snirhizobium spp, Streptococcus spp, Wolinella spp, Xylella spp, Erwinia spp, Agrobacterium spp, Bacillus spp, Paenibacillus spp, Streptomyces spp, Micrococcus spp, Corynebacterium spp, Acetobacter spp, Cyanobacteria spp, Salmonella spp, Rhodococcus spp, Pseudomonas spp, Lactobacillus spp, Enterococcus spp, Alcaligenes spp, Klebsiella spp, Paenibacillus spp, Arthrobacter spp, Corynebacterium spp, Brevibacterium spp, Thermus spp, Pseudomonas spp, Clostridium spp and Es cherichia spp . . Non-limiting examples of bacteria that can be targeted using the methods and compositions provided herein are shown in Table 1.

Table 1. Examples of target bacteria and insect hosts

Endosymbiont Host insect Location 16S rRNA Snodgrassella alvi Honey bee (Apis mellifera) and Bombus spp. Ileum GAGAGTTTGATCCTGGCTCAGATTGAACGCTGGCGGCATGCCTTACACATGCAAGTCGAACGGCAGCACGGAGAGCTTGCTCTCTGGTGGCGAGTGGCGAACGGGTGAGTAATGCATCGGAACGTACCGAGTAATGGGGGATAACTGTCCGAAAGGATGGCTAATACCGCATACGCCCTGAGGGGGAAAGCGGGGGATCGAAAGACCTCGCGTTATTTGAGCGGCCGATGTTGGATTAG CTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCCATAGCGGGTCTGAGAGGATGATCCGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATTTTGGACAATGGGGGGAACCCTGATCCAGCCATGCCGCGTGTCTGAAGAAGGCCTTCGGGTTGTAAAGGACTTTTGTTAGGGAAGAAAAGCCGGGTGTTAATACCATCTGGTGCTGACGTACCTAAAGA ATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTAATCGGAATTACTGGGCGTAAAGCGAGCGCAGACGGTTAATTAAGTCAGATGTGAAATCCCCGAGCTCAACTTGGGACGTGCATTTGAAACTGGTTAACTAGAGTGTGTCAGAGGGAGGTAGAATTCCACGTGTAGCAGTGAAATGCGTAGAGATGTGGAGGAATACCGATGGCGAAGGCAGCCTCCT GGGATAACACTGACGTTCATGCTCGAAAGCGTGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGACAATTAGCTGTTGGGACACTAGATGTCTTAGTAGCGAAGCTAACGCGTGAAATTGTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCAACGCGAAGAACCTTAC CTGGTCTTGACATGTACGGAATCTCTTAGAGATAGGAGAGTGCCTTCGGGAACCGTAACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGAATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCATTAGTTGCCATCATTAAGTTGGGCACTCTAATGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGACCAGGGC TTCACACGTCATACAATGGTCGGTACAGAGGGTAGCGAAGCCGCGAGGTGAAGCCAATCTCAGAAAGCCGATCGTAGTCCGGATTGCACTCTGCAACTCGAGTGCATGAAGTCGGAATCGCTAGTAATCGCAGGTCAGCATACTGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGGGATACCAGAATTGGGTAGACTAACCGCAAGGAGGTCGCTTAAC ACGGTATGCTTCATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTAG (SEQ ID NO: 1) Gilliamella apicola Honey bee (Apis mellifera) and Bombus spp. Ileum TTAAATTGAAGAGTTTGATCATGGCTCAGATTGAACGCTGGCGGCAGGCTTAACACATGCAAGTCGAACGGTAACATGAGTGCTTGCACTTGATGACGAGTGGCGGACGGGTGAGTAAAGTATGGGGATCTGCCGAATGGAGGGGGACAACAGTTGGAAACGACTGCTAATACCGCATAAAGTTGAGAGACCAAAGCATGGGAACCTTCGGGCCATGCGCCATTTGATGAACCCATATGGGATTAG CTAGTTGGTAGGGTAATGGCTTACCAAGGCGACGATCTCTAGCTGGTCTGAGAGGATGACCAGCCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAGCCATGCCGCGTGTATGAAGAAGGCCTTCGGGTTGTAAAGTACTTTCGGTGATGAGGAAGGTGGTGTATCTAATAGGTGCATCAATTGACGTTA ATTACAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCGAGCGTTAATCGGAATGACTGGGCGTAAAGGGCATGTAGGCGGATAATTAAGTTAGGTGTGAAAGCCCTGGGCTCAACCTAGGAATTGCACTTAAAACTGGTTAACTAGAGTATTGTAGAGGAAGGTAGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAATACCGGTGGCGAAGGC GGCCTTCTGGACAGATACTGACGCTGAGATGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCTGTAAACGATGTCGATTTGGAGTTTGTTGCCTAGAGTGATGGGCTCCGAAGCTAACGCGATAAATCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGA AGAACCTTACCTGGTCTTGACATCCACAGAATCTTGCAGAGATGCGGGAGTGCCTTCGGGAACTGTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCCTTTGTTGCCATCGGTTAGGCCGGGAACTCAAAGGAGACTGCCGTTGATAAAGCGGAGGAAGGTGGGGACGACGTCAAGTCATCATGGCCCTTA CGACCAGGGCTACACACGTGCTACAATGGCGTATACAAAGGGAGGCGACCTCGCGAGAGCAAGCGGACCTCATAAAGTACGTCTAAGTCCGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGTGAATCAGAATGTCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCACCAGAAGTAGATAGCTTAACCTTCGGGAG GGCGTTTACCACGGTGTGGTCCATGACTGGGGTGAAGTCGTAACAAGGTAACCGTAGGGGAACCTGCGGTTGGATCACCTCCTTAC (SEQ ID NO: 2) Bartonella apis Honey bee (Apis mellifera) Intestine AAGCCAAAATCAAATTTTCAACTTGAGAGTTTGATCCTGGCTCAGAACGAACGCTGGCGGCAGGCTTAACACATGCAAGTCGAACGCACTTTTCGGAGTGAGTGGCAGACGGGTGAGTAACGCGTGGGAATCTACCTATTTCTACGGAATAACGCAGAGAAATTTGTGCTAATACCGTATACGTCCTTCGGGAGAAAGATTTATCGGAGATAGATGAGCCCGCGTTGGATTAGCTAGTTGGTGA GGTAATGGCCCACCAAGGCGACGATCCATAGCTGGTCTGAGAGGATGACCAGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGCAAGCCTGATCCAGCCATGCCGCGTGAGTGATGAAGGCCCTAGGGTTGTAAAGCTCTTTCACCGGTGAAGATAATGACGGTAACCGGAGAAGAAGCCCCGGCTAACTTCGTGCCAGCAGCCGC GGTAATACGAAGGGGGCTAGCGTTGTTCGGATTTACTGGGCGTAAAGCGCACGTAGGCGGATATTTAAGTCAGGGGTGAAATCCCGGGGCTCAACCCCGGAACTGCCTTTGATACTGGATATCTTGAGTATGGAAGAGGTAAGTGGAATTCCGAGTGTAGAGGTGAAATTCGTAGATATTCGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGTCCATTACTGACGCTGAGGTGCGAAAGCGTG GGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCTGTAAACGATGAATGTTAGCCGTTGGACAGTTTACTGTTCGGTGGCGCAGCTAACGCATTAAACATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGCAGAACCTTACCAGCCCTTGACATCCCGATCGCGGATGGTG GAGACACCGTCTTTCAGTTCGGCTGGATCGGTGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCTTAGTTGCCATCATTTAGTTGGGCACTCTAAGGGGACTGCCGGTGATAAGCCGAGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTACGGGCTGGGCTACACACGTGCTACAATGGTGGTGAC AGTGGGCAGCGAGACCGCGAGGTCGAGCTAATCTCCAAAAGCCATCTCAGTTCGGATTGCACTCTGCAACTCGAGTGCATGAAGTTGGAATCGCTAGTAATCGTGGATCAGCATGCCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTTGGTTTTACCCGAAGGTGCTGTGCTAACCGCAAGGAGGCAGGCAACCACGGTAGGGTCAGCGACTGGGGTGA AGTCGTAACAAGGTAGCCGTAGGGGAACCTGCGGCTGGATCACCTCCTTTCTAAGGAAGATGAAGAATTGGAA (SEQ ID NO: 3) Parasaccharibacter apium Honey bee (Apis mellifera) Intestine CTACCATGCAAGTCGCACGAAACCTTTCGGGGTTAGTGGCGGACGGGTGAGTAACGCGTTAGGAACCTATCTGGAGGTGGGGGATAACATCGGGAAACTGGTGCTAATACCGCATGATGCCTGAGGGCCAAAGGAGAGATCCGCCATTGGAGGGGCCTGCGTTCGATTAGCTAGTTGGTTGGGTAAAGGCTGACCAAGGCGATGATCGATAGCTGGTTTGAGAGGATGATCAGCCACACT GGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGGGCAACCCTGATCCAGCAATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTCACTAGGGAAGATGATGACGGTACCTAGAGAAGAAGCCCCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGGGCTAGCGTTGCTCGGAATGACTGGGCGTAAAGGGCGCGTAGG CTGTTTGTACAGTCAGATGTGAAATCCCCGGGCTTAACCTGGGAACTGCATTTGATACGTGCAGACTAGAGTCCGAGAGAGGGTTGTGGAATTCCCAGTGTAGAGGTGAAATTCGTAGATATTGGGAAGAACACCGGTTGCGAAGGCGGCAACCTGGCTNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN NNNNNNNNNNNNNNNNNNNNNNNNNNNNGAGCTAACGCGTTAAGCACACCGCCTGGGGAGTACGGCCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGCAGAACCTTACCAGGGCTTGCATGGGGAGGCTGTATTCAGAGATGGATATTTCTTCGGACCTCCCGCACAGGTGCTGCATGGCTGTCGTCAGCT CGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCTTTAGTTGCCATCACGTCTGGGTGGGCACTCTAGAGAGACTGCCGGTGACAAGCCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGTCCTGGGCTACACACGTGCTACAATGGCGGTGACAGAGGGATGCTACATGGTGACATGGTGCTGATCTCAAAAAACCGTCTCAGTTCGGATTGT ACTCTGCAACTCGAGTGCATGAAGGTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTTGGTTTGACCTTAAGCCGGTGAGCGAACCGCAAGGAACGCAGCCGACCACCGGTTCGGGTTCAGCGACTGGGGGA (SEQ ID NO: 4) Lactobacillus kunkeei Honey bee (Apis mellifera) Intestine TTCCTTAGAAAGGAGGTGATCCAGCCGCAGGTTCTCCTACGGCTACCTTGTTACGACTTCACCCTAATCATCTGTCCCACCTTAGACGACTAGCTCCTAAAAGGTTACCCCATCGTCTTTGGGTGTTACAAACTCTCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGTGGCATGCTGATCCACGATTACTAGTGATTCCAACTTCATGCAGGCGAGTTGCAGCCTGCA ATCCGAACTGAGAATGGCTTTAAGAGATTAGCTTGACCTCGCGGTTTCGCGACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGCTCATAAGGGGCATGATGATTTGACGTCGTCCCCACCTTTCCTCCGGTTTATCACCGGCAGTCTCACTAGAGTGCCCAACTAAATGCTGGCAACTAATAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAAC CATGCACCACCTGTCATTCTGTCCCCGAAGGGAACGCCCAATCTCTTGGGTTGGCAGAAGATGTCAAGAGCTGGTAAGGTTCTTCGCGTAGCATCGAATTAAACCACATGCTCCACCACTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGAATACTTAATGCGTTAGCTGCGGCACTGAAGGGCGGAAACCCTCCAACACCTAGTATTCATCG TTTACGGCATGGACTACCAGGGTATCTAATCCTGTTCGCTACCCATGCTTTCGAGCCTCAGCGTCAGTAACAGACCAGAAAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTTTCCTCTTCTGTACTCAAGTTTTGTAGTTTCCACTGCACTTCCTCAGTTGAGCTGAGGGCTTTCACAGCAGACTTACAAAACCGCCTGCGCTCGCTTTACGCC CAATAAATCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAAATACCGTCAAAGTGTTAACAGTTACTCTAACACTTGTTCTTCTTTAACAACAGAGTTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCATCAGACTTTCGTCCATTGTGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTC AGTCCCAATGTGGCCGATTACCCTCTCAGGTCGGCTACGTATCATCGTCTTGGTGGGCTTTTATCTCACCAACTAACTAATACGGCGCGGGTCCATCCCAAAGTGATAGCAAAGCCATCTTTCAAGTTGGAACCATGCGGTTCCAACTAATTATGCGGTATTAGCACTTGTTTCCAAATGTTATCCCCCGCTTCGGGGCAGGTTACCCACGTGTTACTCACCAGTTCGCCACTCGCTCCGAATCCAAAA ATCATTTATGCAAGCATAAAATCAATTTGGGAGAACTCGTTCGACTTGCATGTATTAGGCACGCCGCCAGCGTTCGTCCTGAGCCAGGATCAAACTCTCATCTTAA (SEQ ID NO: 190) Lactobacillus Firm-4 Honey bee (Apis mellifera) Intestine ACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAGCGCGGGAAGTCAGGGAAGCCTTCGGGTGGAACTGGTGGAACGAGCGGCGGATGGGTGAGTAACACGTAGGTAACCTGCCCTAAAGCGGGGGATACCATCTGGAAACAGGTGCTAATACCGCATAAACCCAGCAGTCACATGAGTGCTGGTTGAAAGACGGCTTCGGCTGTCACTTTAGGATGGACCTGCGGCGTATTAGCTAGTTGG TGGAGTAACGGTTCACCAAGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGGATGAAGAAGGTCTTCGGATCGTAAAATCCTGTTGTTGAAGAAGAACGGTTGTGAGAGTAACTGCTCATAACGTGACGGTAATCAACCAG AAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGGGAGCGCAGGCGGTCTTTTTAAGTCTGAATGTGAAAGCCCTCAGCTTAACTGAGGAAGAGCATCGGAAACTGAGAGACTTGAGTGCAGAAGAGGAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTCT CTGGTCTGTTACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGGAGGTTTCCGCCTCTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACC TTACCAGGTCTTGACATCTCCTGCAAGCCTAAGAGATTAGGGGTTCCCTTCGGGGACAGGAAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTACTAGTTGCCAGCATTAAGTTGGGCACTCTAGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGACGACGTCAAATCATCATGCCCCTTATGACCTGGGC TACACACGTGCTACAATGGATGGTACAATGAGAAGCGAACTCGCGAGGGGAAGCTGATCTCTGAAAACCATTCTCAGTTCGGATTGCAGGCTGCAACTCGCCTGCATGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCC (SEQ ID NO: 191) Enterococcus Bombyx mori Intestine AGGTGATCCAGCCGCACCTTCCGATACGGCTACCTTGTTACGACTTCACCCCAATCATCTATCCCACCTTAGGCGGCTGGCTCCAAAAAGGTTACCTCACCGACTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCGTGCTGATCCGCGATTACTAGCGATTCCGGCTTCATGCAGGCGAGTTGCAGCCTGCAATCCGAACTGAGA GAAGCTTTAAGAGATTTGCATGACCTCGCGGTCTAGCGACTCGTTGTACTTCCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCGTAGAGTGCCCAACTAAATGATGGCAACTAACAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTG TCACTTTGTCCCCCGAAGGGAAAGCTCTATCTCTAGAGTGGTCAAAGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGAGTGCTTAATGCGTTTGCTGCAGCACTGAAGGGCGGAAACCCTCAACACTTAGCACTCATCGTTTACGGCGTG GACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGAGCCTCAGCGTCAGTTACAGACCAGAGAGCCGCCTTCGCCACTGGTGTTCCTCCATATATCTACGCATTTCACCGCTACACATGGAATTCCACTCTCCTCTTCTGCACTCAAGTCTCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGACA ACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGATACCGTCAGGGGACGTTCAGTTACTAACGTCCTTGTTCTTCTCTAACAACAGAGTTTTACGATCCGAAAACCTTCTTCACTCACGCGGCGTTGCTCGGTCAGACTTTCGTCCATTGCCGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGC CGATCACCCTCTCAGGTCGGCTATGCATCGTGGCCTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCACCGCGGGTCCATCCATCAGCGACACCCGAAAGCGCCTTTCACTCTTATGCCATGCGGCATAAACTGTTATGCGGTATTAGCACCTGTTTCCAAGTGTTATCCCCCTCTGATGGGTAGGTTACCCACGTGTTACTCACCCGTCCGCCACTCCTCTTTCCAATTGAGTGCAAGCACTCG GGAGGAAAGAAGCGTTCGACTTGCATGTATTAGGCACGCCGCCAGCGTTCGTCCTGAGCCAGGATCAAACTCT (SEQ ID NO: 5) Delftia Bombyx mori Intestine CAGAAAGGAGGTGATCCAGCCGCACCTTCCGATACGGCTACCTTGTTACGACTTCACCCCAGTCACGAACCCCGCCGTGGTAAGCGCCCTCCTTGCGGTTAGGCTACCTACTTCTGGCGAGACCCGCTCCCATGGTGTGACGGGCGGTGTGTACAAGACCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCACGCAGTCGAGTTGCAGACTGCGATCCG GACTACGACTGGTTTTATGGGATTAGCTCCCCCTCGCGGGTTGGCAACCCTCTGTACCAGCCATTGTATGACGTGTGTAGCCCCACCTATAAGGGCCATGAGGACTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCATTAGAGTGCTCAACTGAATGTAGCAACTAATGACAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAGC CATGCAGCACCTGTGCAGGTTCTCTTTCGAGCACGAATCCATCTCTGGAAACTTCCTGCCATGTCAAAGGTGGGTAAGGTTTTTCGCGTTGCATCGAATTAAACCACATCATCCACCGCTTGTGCGGGTCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGCCGTACTCCCCAGGCGGTCAACTTCACGCGTTAGCTTCGTTACTGAGAAAACTAATTCCCAACAACCAGTTGACATCGTTTAGGGC GTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGTGCATGAGCGTCAGTACAGGTCCAGGGGATTGCCTTCGCCATCGGTGTTCCTCCGCATATCTACGCATTTCACTGCTACACGCGGAATTCCATCCCCCTCTACCGTACTCTAGCCATGCAGTCACAAATGCAGTTCCCAGGTTGAGCCCGGGGATTTCACATCTGTCTTACATAACCGCCTGCGCACGCTTTACGCCCAG TAATTCCGATTAACGCTCGCACCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGTGCTTATTCTTACGGTACCGTCATGGGCCCCCTGTATTAGAAGGAGCTTTTTCGTTCCGTACAAAAGCAGTTTACAACCCGAAGGCCTTCATCCTGCACGCGGCATTGCTGGATCAGGCTTTCGCCCATTGTCCAAAATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTC CCAGTGTGGCTGGTCGTCCTCTCAGACCAGCTACAGATCGTCGGCTTGGTAAGCTTTTATCCCACCAACTACCTAATCTGCCATCGGCCGCTCCAATCGCGCGAGGCCCGAAGGGCCCCCGCTTTCATCCTCAGATCGTATGCGGTATTAGCTACTCTTTCGAGTAGTTATCCCCCACGACTGGGCACGTTCCGATGTATTACTCACCCGTTCGCCACTCGTCAGCGTCCGAAGACCTGTTACCG TTCGACTTGCATGTGTAAGGCATGCCGCCAGCGTTCAATCTGAGCCAGGATCAAACTCTACAGTTCGATCT (SEQ ID NO: 6) Pelomonas Bombyx mori Intestine ATCCTGGCTCAGATTGAACGCTGGCGGCATGCCTTACACATGCAAGTCGAACGGTAACAGGTTAAGCTGACGAGTGGCGAACGGGTGAGTAATATATCGGAACGTGCCCAGTCGTGGGGGATAACTGCTCGAAAGAGCAGCTAATACCGCATACGACCTGAGGGTGAAAGCGGGGGATCGCAAGACCTCGCNNGATTGGAGCGGCCGATATCAGATTAGGTAGTTGGTGGGGTAAAGGCCCA CCAAGCCAACGATCTGTAGCTGGTCTGAGAGGACGACCAGCCACACTGGGACTGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATTTTGGACAATGGGCGCAAGCCTGATCCAGCCATGCCGCGTGCGGGAAGAAGGCCTTCGGGTTGTAAACCGCTTTTGTCAGGGAAGAAAAGGTTCTGGTTAATACCTGGGACTCATGACGGTACCTGAAGAATAAGCACCGGCTAACTACG TGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTTATGCAAGACAGAGGTGAAATCCCCGGGCTCAACCTGGGAACTGCCTTTGTGACTGCATAGCTAGAGTACGGTAGAGGGGGATGGAATTCCGCGTGTAGCAGTGAAATGCGTAGATATGCGGAGGAACACCGATGGCGAAGGCAATCCCCTGGACCTGTACTGACGCTCAT GCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGTCAACTGGTTGTTGGGAGGGTTTCTTCTCAGTAACGTANNTAACGCGTGAAGTTGACCGCCTGGGGAGTACGGCCGCAAGGTTGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGTTTAATTCGATGCAACGCGAAAAACCTTACCTACCCTTGACAT GCCAGGAATCCTGAAGAGATTTGGGAGTGCTCGAAAGAGAACCTGGACACAGGTGCTGCATGGCCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCATTAGTTGCTACGAAAGGGCACTCTAATGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAGGTCATCATGGCCCTTATGGGTAGGGCTACACACGTCATACAAT GGCCGGGACAGAGGGCTGCCAACCCGCGAGGGGGAGCTAATCCCAGAAACCCGGTCGTAGTCCGGATCGTAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGCGGATCAGCTTGCCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGCGGGTTCTGCCAGAAGTAGTTAGCCTAACCGCAAGGAGGGCGATTACCACGGCAGGGTT CGTGACTGGGGTGAAGTCGTAACAAGGTAGCCGTATCGGAAGGTGCGGCTGGATCAC (SEQ ID NO: 7)

Any number of bacterial species can be targeted using the compositions or methods described herein. For example, in some cases the modulating agent may specifically target one type of bacteria. In some cases, the modulating agent may specifically target at least about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80 , 90, 100, 150, 200, 250, 500 or more different types of bacteria. In some cases, the modulating agent may target any of about 1 to about 5, about 5 to about 10, about 10 to about 20, about 20 to about 50, about 50 to about 100, about 100 to about 200, about 200 to about 500, about 10 to about 50, about 5 to about 20, or about 10 to about 100 different species of bacteria. In some cases, the modulating agent may specifically target at least about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80 , 90, 100 or more types, classes, orders, families or genera of bacteria.

In some cases, the modulating agent may increase the population of one or more bacteria (e.g., commensal bacteria) by at least approximately any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater in the host compared to a host to which the modulating agent was not administered. In some cases, the modulating agent may reduce the population of one or more bacteria (eg, pathogenic or parasitic bacteria) by at least approximately any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80 %, 90% or more in the host compared to a host to which the modulating agent was not administered. In some cases, the modulating agent can destroy a population of bacteria (eg, pathogenic or parasitic bacteria) in the host. In some cases, the modulating agent may increase the level of one or more commensal bacteria by at least about any value of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in host and/or can reduce the level of one or more pathogenic bacteria by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in host compared to a host to which the modulating agent was not administered.

In some cases, the modulating agent may alter bacterial diversity and/or bacterial composition of the host. In some cases, the modulating agent may increase bacterial diversity in the host relative to baseline diversity by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to a host to which the modulating agent was not administered. In some cases, the modulating agent may reduce the diversity of bacteria in the host relative to the baseline diversity by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to a host to which the modulating agent was not administered.

In some cases, the modulating agent may alter the function, activity, growth and/or division of one or more bacterial cells. For example, a modulating agent may alter the expression of one or more genes in a bacterium. In some cases, the modulating agent may change the function of one or more proteins in the bacterium. In some cases, the modulating agent may alter the function of one or more cellular structures (eg, cell wall, outer or inner membrane) in the bacterium. In some cases, the modulating agent can destroy (eg, lyse) the bacterium.

The target bacterium may live in one or more parts of the insect's body. In addition, the target bacterium may be intracellular or extracellular. In some cases, the bacteria reside in one or more parts of the host's gut, including, for example, the foregut, midgut, and/or hindgut. For example, in honey bees (e.g. Apis mellifera ), bacterial symbionts confined to the adult hindgut are acquired in the first few days after adults emerge from the pupal stage through social interactions with other adult worker bees in the colony. The inhabitants of the honey bee gut belong to a small number of characteristic lineages found only in honey bees, as well as in other Apis species and in Bombus species (bumblebees). In some cases, the target bacteria live in the honey bee's body. In some cases, one or more of the target bacteria in the honey bee is Snodgrassella spp. (e.g. Snodgrassella alvi ), Gilliamella spp. (eg Gilliamella apicola ), Bartonella spp. (eg Bartonella apis ), Parasaccharibacter spp. (eg Parasaccharibacter apium ) or Lactobacillus spp. (e.g. Lactobacillus kunkeei , Lactobacillus Firm-4 ).

In some cases, the bacteria live as intracellular bacteria in the host insect cell. In some cases, bacteria live in the bacteriocyte of the host insect.

Changes in bacterial populations in the host can be determined using any methods known in the art, such as microarray analysis, polymerase chain reaction (PCR), real-time PCR, flow cytometry, fluorescence microscopy, transmission electron microscopy, fluorescence in situ hybridization (eg, FISH), spectrophotometry, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), and DNA sequencing. In some cases, a sample from a host treated with a modulating agent is sequenced (eg, by metagenomic sequencing of 16S rRNA or rDNA) to determine the host microbiota following delivery or administration of the modulating agent. In some cases, a sample from a host that has not received the modulating agent is also sequenced to provide a reference.

ii. Mushrooms

Illustrative fungi that may be targeted in accordance with the methods and compositions provided herein include, but are not limited to, Amylostereum areolatum, Epichloe spp, Pichia pinus, Hansenula capsulate, Daldinia decipien, Ceratocytis spp, Ophiostoma spp, and Attamyces bromatificus. Non-limiting examples of yeast and yeast-like symbionts found in insects include Candida, Metschnikowia, Debaryomyces, Scheffersomyces shehatae and Scheffersomyces stipites, Starmerella, Pichia, Trichosporon, Cryptococcus, Pseudozyma, as well as yeast-like symbionts of the subphylum Pezizomycotina ( for example Symbiotaphrina bucneri and Symbiotaphrina kochii ). Non-limiting examples of yeasts that can be targeted using the methods and compositions herein are shown in Table 2.

Table 2. Examples of yeast fungi in insects

Insect type Squad: family Location of yeast fungi (species) Stegobium paniceum Coleoptera: Anobiidae Mycetomas (= Sitodrepa panicea) (Saccharomyces) Cecum (Torulopsis buchnerii) Mycetoma between the foregut and midgut Mycetomas (Symbiotaphrina buchnerii) Mycetomas and digestive tube (Torulopsis buchnerii) Cecum (Symbiotaphrina buchnerii) Lasioderma serricorne Coleoptera: Anobiidae Mycetoma between the foregut and midgut (Symbiotaphrina kochii) Ernobius abietis Coleoptera: Anobiidae Mycetomas (Torulopsis karawaiewii) (Candida karawaiwii) Ernobius mollis Coleoptera: Anobiidae Mycetomas (Torulopsis ernobii) (Candida ernobii) Hemicoelus gibbicollis Coleoptera: Anobiidae Mycetomas of larvae Xestobium plumbeum Coleoptera: Anobiidae Mycetomas (Torulopsis xestobii) (Candida xestobii) Criocephalus rusticus Coleoptera: Cerambycidae Mycetomas Phoracantha semipunctata Coleoptera: Cerambycidae Alimentary canal (Candida guilliermondii, C. tenuis) Caecum near the midgut (Candida guilliermondii) Harpium inquisitor Coleoptera: Cerambycidae Mycetomas (Candida rhagii) Harpium mordax
H. sycophanta Coleoptera: Cerambycidae Caecum near the midgut (Candida tenuis) Gaurotes virginea Coleoptera: Cerambycidae Cecum near midgut (Candida rhagii) Leptura rubra Coleoptera: Cerambycidae Caecum near the midgut (Candida tenuis) Cecum near the midgut (Candida parapsilosis) Leptura maculicornis Coleoptera: Cerambycidae Cecum near the midgut (Candida parapsilosis) L. cerambyciformis Leptura sanguinolenta Coleoptera: Cerambycidae Cecum near midgut (Candida sp.) Rhagium bifasciatum Coleoptera: Cerambycidae Caecum near the midgut (Candida tenuis) Rhagium inquisitor Coleoptera: Cerambycidae Caecum near the midgut (Candida guilliermondii) Rhagium mordax Coleoptera: Cerambycidae Caecum near the midgut (Candida) Carpophilus hemipterus Coleoptera: Nitidulidae Intestinal tract (10 types of yeasts) Odontotaenius disjunctus Coleoptera: Passalidae Hindgut (Enteroramus dimorphus) Odontotaenius disjunctus Coleoptera: Passalidae Intestine (Pichia stipitis, P. segobiensis, Candida shehatae) Verres sternbergianus (C. ergatensis) Scarabaeus semipunctatus Coleoptera: Scarabaeidae Digestive tract (10 types of yeasts) Chironitis furcifer Unknown species Coleoptera: Scarabaeidae Gut (Trichosporon cutaneum) Dendroctonus and Ips spp. Coleoptera: Scolytidae Alimentary canal (13 types of yeast) Dendroctonus frontalis Coleoptera: Scolytidae Midgut (Candida sp.) Ips sexdentatus Coleoptera: Scolytidae Digestive tract (Pichia bovis, P. rhodanensis) Hansenula holstii (Candida rhagii) Digestive tract (Candida pulcherina) Ips typographus Coleoptera: Scolytidae Alimentary canal Digestive tract (Hansenula capsulata, Candida parapsilosis) Gut and homogenates of beetles (Hansenula holstii, H. capsulata, Candida diddensii, C. mohschtana, C. nitratophila, Cryptococcus albidus, C. laurentii) Trypodendron lineatum Coleoptera: Scolytidae Undefined Xyloterinus politus Coleoptera: Scolytidae Head, chest, abdomen (Candida, Pichia, Saccharomycopsis) Periplaneta americana Dictyoptera: Blattidae Hemocoel (Candida sp. nov.) Blatta orientalis Dictyoptera: Blattidae Intestinal tract (Kluyveromyces blattae) Blatella germanica Dictyoptera: Blattellidae Hemocoel Cryptocercus punctulatus Dictyoptera: Cryptocercidae Hindgut (1 species of yeast) Philophylla heraclei Diptera: Tephritidae Hemocoel Aedes (4 species) Diptera: Culicidae Internal microflora (9 genera of yeast fungi) Drosophila pseudoobscura Diptera: Drosophilidae Alimentary canal (24 types of yeast) Drosophila (5 species) Diptera: Drosophilidae Goiter (42 types of yeasts) Drosophila melanogaster Diptera: Drosophilidae Goiter (8 types of yeast) Drosophila (4 species) Diptera: Drosophilidae Goiter (7 types of yeast) Drosophila (6 species) Diptera: Drosophilidae Gut of larvae (17 species of yeasts) Drosophila (20 species) Diptera: Drosophilidae Goiter (20 types of yeast) Drosophila (8 species groups) Diptera: Drosophilidae Goiter (Kloeckera, Candida, Kluyveromyces) Drosophila serido Diptera: Drosophilidae Goiter (18 species of yeast) Drosophila (6 species) Diptera: Drosophilidae Intestinal epithelium (Coccidiascus legeri) Protaxymia melanoptera Diptera Unknown (Candida, Cryptococcus, Sporoblomyces) Astegopteryx styraci Homoptera: Aphididae Hemocoel and fat body Tuberaphis sp. Homoptera: Aphididae Tissue sections Hamiltonaphis styraci Glyphinaphis bambusae Cerataphis sp. Hamiltonaphis styraci Homoptera: Aphididae Abdominal hemocoel Cofana unimaculata Homoptera: Cicadellidae Fat body Leofa unicolor Homoptera: Cicadellidae Fat body Lecaniines, etc. Homoptera: Coccoidea Hemolymph, adipose tissue, etc. Lecanium sp. Homoptera: Coccidae Hemolymph, adipose tissue Ceroplastes (4 species) Homoptera: Coccidae Blood smears Laodelphax striatellus Homoptera: Delphacidae Fat body Eggs Eggs (Candida) Nilaparvata lugens Homoptera: Delphacidae Fat body Eggs (2 unidentified yeast species) Eggs, nymphs (Candida) Eggs (7 unidentified yeast species) Eggs (Candida) Nisia nervosa Homoptera: Delphacidae Fat body Nisia grandiceps Perkinsiella spp. Sardia rostrata Sogatella furcifera Sogatodes orizicola Homoptera: Delphacidae Fat body Amrasca devastans Homoptera: Jassidae Eggs, mycetomas, hemolymph Tachardina lobata Homoptera: Kerriidae Blood smears (Torulopsis) Laccifer (=Lakshadia) sp. Homoptera: Kerriidae Blood smears (Torula variabilis) Comperia merceti Hymenoptera: Encyrtidae Hemolymph, intestine, poison gland Solenopsis invicta Hymenoptera: Formicidae Hemolymph (Myrmecomyces annellisae) S. quinquecuspis Solenopsis invicta Hymenoptera: Formicidae Larvae at the fourth instar stage (Candida parapsilosis, Yarrowia lipolytica) Gut and hemolymph (Candida parapsilosis, C. lipolytica, C. guillermondii, C. rugosa, Debaryomyces hansenii) Apis mellifera Hymenoptera: Apidae Digestive tract (Torulopsis sp.) Intestinal tract (Torulopsis apicola) Digestive tract (8 types of yeasts) Gut contents (12 types of yeast) Intestinal contents (7 types of yeast) Intestines (14 types of yeasts) Intestinal tract (Pichia melissophila) Intestinal tract (7 types of yeasts) Alimentary canal (Hansenula silvicola) Crop and gut (13 types of yeasts) Apis mellifera Hymenoptera: Apidae Midgut (9 genera of yeasts) Anthophora occidentalis Hymenoptera: Anthophoridae Nomia melanderi Hymenoptera: Halictidae Halictus rubicundus Hymenoptera: Halictidae Megachile rotundata Hymenoptera: Megachilidae

Any number of fungal species can be targeted using the compositions or methods described herein. For example, in some cases the modulating agent may specifically target one type of fungus. In some cases, the modulating agent may specifically target at least about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80 , 90, 100, 150, 200, 250, 500 or more different types of mushrooms. In some cases, the modulating agent may target any of about 1 to about 5, about 5 to about 10, about 10 to about 20, about 20 to about 50, about 50 to about 100, about 100 to about 200, from about 200 to about 500, from about 10 to about 50, from about 5 to about 20, or from about 10 to about 100 different types of mushrooms. In some cases, the modulating agent may specifically target at least about any of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80 , 90, 100 or more types, classes, orders, families or genera of fungi.

In some cases, the modulating agent may increase the population of one or more fungi (e.g., symbiotic fungi) by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater in the host compared to a host to which the modulating agent was not administered. In some cases, the modulating agent may reduce the population of one or more fungi (eg, pathogenic or parasitic fungi) by at least approximately any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80 %, 90% or more in the host compared to a host to which the modulating agent was not administered. In some cases, the modulating agent can destroy the fungal population (eg, pathogenic or parasitic fungi) in the host. In some cases, the modulating agent may increase the level of one or more symbiotic fungi by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in host and/or can reduce the level of one or more pathogenic fungi by at least approximately any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more in host compared to a host to which the modulating agent was not administered.

In some cases, the modulating agent may alter the fungal diversity and/or fungal composition of the host. In some cases, the modulating agent may increase fungal diversity in the host relative to baseline diversity by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to a host to which the modulating agent was not administered. In some cases, the modulating agent may reduce the diversity of fungi in the host relative to baseline diversity by at least about any of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more compared to a host to which the modulating agent was not administered.

In some cases, the modulating agent may alter the function, activity, growth and/or division of one or more fungi. For example, a modulating agent may alter the expression of one or more genes in a fungus. In some cases, the modulating agent may change the function of one or more proteins in the fungus. In some cases, the modulating agent may alter the function of one or more cellular components in the fungus. In some cases, the modulating agent can destroy the fungus.

In addition, the target fungus may live in one or more parts of the insect's body. In some cases, the fungus resides in or on one or more parts of the insect's gut, including, for example, the foregut, midgut, and/or hindgut. In some cases, the fungus lives extracellularly in the hemolymph, fat bodies, or specialized structures in the host.

Changes in the fungal population in the host can be determined using any methods known in the art, such as microarray analysis, polymerase chain reaction (PCR), real-time PCR, flow cytometry, fluorescence microscopy, transmission electron microscopy, fluorescence in situ hybridization (eg, FISH), spectrophotometry, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), and DNA sequencing. In some cases, a sample from a host treated with a modulating agent is sequenced (eg, by metagenomic sequencing) to determine the host microbiota following delivery or administration of the modulating agent. In some cases, a sample from a host that has not received the modulating agent is also sequenced to provide a reference.

III. Modulating agents

The modulating agent of the methods and compositions provided herein may include a phage, polypeptide, small molecule, antibiotic, secondary metabolite, bacterium, or any combination thereof.

i. Phage

The modulating agent described herein may include a phage (eg, a lytic phage or a non-lytic phage). In some cases, an effective concentration of any phage described herein may alter the level, activity, or metabolism of one or more microorganisms (described herein) resident in a host described herein, the change resulting in increased fitness of the host. In some cases, the modulating agent includes at least one phage selected from the orders Tectiviridae, Myoviridae, Siphoviridae, Podoviridae, Caudovirales, Lipothrixviridae, Rudiviridae or Ligamenvirales . In some cases , the composition contains at least one phage selected from the family Myoviridae, Siphoviridae, Podoviridae, Lipothrixviridae, Rudiviridae, Ampullaviridae, Bicaudaviridae, Clavaviridae, Corticoviridae, Cystoviridae, Fuselloviridae, Gluboloviridae, Guttaviridae, Inoviridae, Leviviridae, Microviridae, Plasmaviridae and Tectiviridae . Additional non-limiting examples of phages useful in the methods and compositions are provided in Table 3.

Table 3. Examples of phages and target bacteria

Phage and accession no. Target bacterium Host-target SA1(NC_027991), phiP68 (NC_004679) Staphylococcus sp. Family Apidae WO (AB036666.1) Wolbachia sp. Aedes aegypti; Drosophila melanogaster; Plasmodium sp; Teleogryllus taiwanemma; Bombyx mori KL1 (NC_018278), BcepNazgul (NC_005091) PhiE125 (NC_003309) Burkholderia sp. Riptortus sp.; Pyrrhocoris apterus. Fern (NC_028851), Xenia (NC_028837), HB10c2 (NC_028758) Paenibacillus larvae Bumblebees: Bombus sp.; honey bees: A. mellifera CP2 (NC_020205), XP10 (NC_004902), XP15 (NC_007024), phiL7 (NC_012742) Xanthomonas sp. Plebeina denoiti; family Apidae; Apis mellifera; family Drosophilidae and family Chloropidae PP1 (NC_019542), PM1 (NC_023865) Pectobacterium carotovorum subsp. carotovorum Family Apidae ΦRSA1 (NC_009382),
ΦRSB1 (NC_011201), ΦRSL1 (NC_010811), RSM1 (NC_008574) Ralstonia solanacearum Bombyx mori SF1(NC_028807) Streptomyces scabies Philanthus sp.; Trachypus sp. ECML-4 (NC_025446), ECML-117 (NC_025441), ECML-134 (NC_025449) Escherichia coli Family Apidae; Varroa destructor SSP5(JX274646,1), SSP6 (NC_004831), SFP10 (NC_016073), F18SE (NC_028698) Salmonella sp. Family Drosophilidae γ (NC_001416), Bcp1 (NC_024137) Bacillus sp. Gypsy moth; Lymantria dispar; Varroa destructor Phi1 (NC_009821) Enterococcus sp. Schistocerca gragaria ΦKMV (NC_005045), ΦEL(AJ697969,1), ΦKZ (NC_004629) Pseudomonas sp. Lymantria dispar; family Apidae A2 (NC_004112), phig1e (NC_004305) Lactobacillus sp. Family Apidae; family Drosophila; Varroa destructor KLPN1 (NC_028760) Klebsiella sp. C. capitata vB_AbaM_Acibel004 (NC_025462), vB_AbaP_Acibel007 (NC_025457) Acinetobacter sp. Schistocerca gragaria

In some cases, the modulating agent includes a lytic phage. Thus, after delivering a lytic phage to a bacterial cell residing in the host, the phage causes lysis in the target bacterial cell. In some cases, a lytic phage specifically attacks a bacterium living in the body of an insect host and destroys it, increasing the fitness of the host. Alternatively or additionally, the modulating agent phage may be a non-lytic phage (also called a lysogenic or temperate phage). Thus, after delivery of a non-lytic phage to a bacterial cell living in the host body, the bacterial cell can remain viable and capable of stably maintaining the expression of genes encoded in the phage genome. In some cases, a non-lytic phage is used to alter gene expression in a bacterium living in an insect host, increasing the fitness of the host. In some cases, the modulating agent includes a mixture of lytic and non-lytic phage.

The modulating agent described herein may include a phage with a narrow or broad spectrum of target bacteria. In some cases, the phage has a narrow range of target bacteria. In some cases, the phage is a promiscuous phage with a wide range of target bacteria. For example, a promiscuous phage can specifically target at least about any of the 5, 10, 20, 30, 40, 50 or more bacteria present in the host. A phage with a narrow spectrum of target bacteria can specifically affect a specific strain of bacteria in the host without negatively affecting another, such as non-target, bacterium in the host. For example, a phage can target no more than about 50, 40, 30, 20, 10, 8, 6, 4, 2, or 1 bacteria found in the host. For example, the compositions described herein can target the bacterial pathogen Paenibacillus larvae in honey bees, but cannot target other bacteria (eg, commensal bacteria), including Lactobacillus Firm4 , Lactobacillus Firm5 , Bifidobacterium sp, Snodgrassella alvi , Gilliamella apicola , Bartonella apis , Parasaccharibacter apium or Lactobacillus kunkeei . In some cases, a phage infects one or more specific bacteria by binding to a cell surface protein (e.g. OmpF, LamB, BtuB, TolC, etc.) or another recognition molecule (e.g. glycolipids, LPS, lipoteichoic acid, etc. .).

The compositions described herein may contain any number of phages, such as at least about any number of 1, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 100 or more phages . In some cases, the composition contains a phage from one or more families (for example, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more phages), one or more orders (for example, 2, 3, 4, 5 , 6, 7, 8, 9, 10 or more phages) or one or more species (for example, 1, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 100 or more phages). Compositions containing one or more phages are also referred to herein as “phage cocktails.” Phage cocktails are useful because they provide targeted targeting to a wider range of bacterial hosts. In addition, they provide targeted exposure to each strain (i.e., subspecies) of bacteria of numerous orthogonal phages, thereby preventing or significantly delaying the emergence of resistance. In some cases, the cocktail contains several phages that specifically target one type of bacteria. In some cases, the cocktail contains several phages that specifically target several types of bacteria. In some cases, a single-phage “cocktail” contains one promiscuous phage (i.e., a phage with a wide host range) that specifically targets many strains within a species.

Suitable concentrations of phage in the modulating agent described herein depend on factors such as potency, survival rate, phage tolerability, the number of different phages and/or types of lysine in the compositions, composition and methods of application of the composition. In some cases, the phage is in a liquid or solid composition. In some cases, the concentration of each phage in any of the compositions described herein is at least approximately any of 10 ² , 10 ³ , 10 4 , 10 ⁵ , 10 ⁶ , 10 ⁷ , ^{10 8 ,} 10 ⁹ , 10 ¹⁰ or more PFU/ml. In some cases, the concentration of each phage in any of the compositions described herein is no more than approximately any value of 10 ² , 10 ³ , 10 4 , 10 ⁵ , 10 ⁶ , 10 ⁷ , ^{10 8} , 10 ⁹ PFU/ml . In some cases, the concentration of each phage in the composition is any of from about 10 ² to about 10 ³ , from about 10 ³ to about 10 ⁴ , from about 10 ⁴ to about 10 ⁵ , from about 10 ⁵ to about 10 ⁶ , from about 10 ⁷ to about 10 ⁸ , from about 10 ⁸ to about 10 ⁹ , from about 10 ² to about 10 ⁴ , from about 10 ⁴ to about 10 ⁶ , from about 10 ⁶ to about 10 ⁹ , or from about 10 ³ to about 10 ⁸ PFU/ml. In some cases, if the composition contains at least two types of phages, the concentration of each type of phages may be the same or different. For example, in some cases, the concentration of one phage in the cocktail is approximately 10 ⁸ PFU/ml, and the concentration of a second phage in the cocktail is approximately 10 ⁶ PFU/ml.

The modulating agent, including a phage described herein, can be brought into contact with the target host in an amount and for a time sufficient to: (a) achieve a target level (for example, a predetermined or threshold level) concentration of the phage within target host; (b) achieving a target level (eg, a predetermined or threshold level) of phage concentration within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of phage concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/and (e) modulating the fitness of the target host.

As illustrated in Examples 7 and 11, phages can be used as modulating agents in the elimination of bacterial pathogens such as Serratia marcescens, Erwinia carotovora and Pseudomonas entomophila from insect hosts such as honey bees or silkworms.

ii. Polypeptides

Numerous polypeptides (eg, bacteriocin, R-type bacteriocin, C-rich nodule peptide, antimicrobial peptide, lysine, or bacteriocytic regulatory peptide) can be used in the compositions and methods described herein. In some cases, an effective concentration of any peptide or polypeptide described herein can alter the level, activity or metabolism of one or more microorganisms (described above) resident in the host, the change resulting in increased fitness of the host. Polypeptides included herein may include naturally occurring polypeptides or recombinantly produced variants. For example, the polypeptide may be a functionally active variant of any of the polypeptides described herein that is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79 %, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical, for example, in a specific region or in the entire sequence, to the sequence of a polypeptide described herein or a naturally occurring polypeptide.

A modulatory agent comprising a polypeptide described herein may be brought into contact with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) concentration within the host -targets; (b) achieving a target level (eg, a predetermined or threshold level) of concentration within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/and (e) modulating the fitness of the target host.

Polypeptide modulatory agents discussed later in this document, namely bacteriocins, lysines, antimicrobial peptides, C-rich nodule peptides and bacteriocyte regulatory peptides, can be used to alter the level, activity or metabolism of target microorganisms, as discussed in the section on enhancing insect fitness , such as honey bees and silkworms.

(a) Bacteriocins

The modulating agent described herein may include a bacteriocin. In some cases, the bacteriocin is produced naturally by gram-positive bacteria such as Pseudomonas, Streptomyces, Bacillus, Staphylococcus or lactic acid bacteria (LAB, such as Lactococcus lactis ). In some cases, the bacteriocin is produced naturally by gram-negative bacteria such as Hafnia alvei , Citrobacter freundii , Klebsiella oxytoca, Klebsiella pneumonia, Enterobacter cloacae , Serratia plymithicum, Xanthomonas campestris, Erwinia carotovora, Ralstonia solanacearum or Escherichia coli . Exemplary bacteriocins include, but are not limited to, LAB class I-IV antibiotics (such as lantibiotics), colicins, microcins, and pyocins. Non-limiting examples of bacteriocins are shown in Table 4.

Table 4. Examples of bacteriocins

Class Name Producer Targets Subsequence Class I Nizin Lactococcus lactis Active against gram-positive bacteria: Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Listeria, Clostridium ITSISLCTPGCKTGALMGCNMKTATCHCSIHVSK (SEQ ID NO: 8) Epidermin Staphylococcus epidermis Gram-positive bacteria IASKFICTPGCAKTGSFNSYCC (SEQ ID NO: 9) Class II Class II a Pediocin PA-1 Pediococcus acidilactici Pediococci, Lactobacilli, Leuconostoc, Brochothrix thermosphacta, Propionibacteria, Bacilli, Enterococci, Staphylococci, Listeria clostridia, Listeria monocytogenes, Listeria innocua KYYGNGVTCG KHSCSVDWGK ATTCIINNGAMAWATGGHQGNHKC (SEQ ID NO: 10) Class II b Enterocin P Enterococcus faecium Lactobacillus sakei, Enterococcus faecium ATRSYGNGVYCNNSKCWVNWGEAKENIAGIVISGWASGLAGMGH (SEQ ID NO: 11) Class IIc Lactococcin G Streptococcus lactis Gram-positive bacteria GTWDDIGQGIGRVAYWVGKAMGNMSDVNQASRINRKKKH (SEQ ID NO: 12) Class II d Lactacin-F Lactobacillus johnsonii Lactobacilli, Enterococcus faecalis NRWGDTVLSAASGAGTGIKACKSFGPWGMAICGVGGAAIGGYFGYTHN (SEQ ID NO: 13) Class III Class IIIa Enterocin AS-48 Enterococcus faecalis Broad spectrum: gram-positive and gram-negative bacteria MAKEFGIPAAVAGTVLNVVEAGGWVTTIVSILTAVGSGGLSLLAAAGRESIKAYLKKEIKKKGKRAVIAW (SEQ ID NO: 14) Class III b Aureocin A70 Staphylococcus aureus Broad spectrum: gram-positive and gram-negative bacteria MSWLNFLKYIAKYGKKAVSAAWKYKGKVLEWLNVGPTLEWVWQKLKKIAGL (SEQ ID NO: 15) Class IV Garvicin A Lactococcus garvieae Broad spectrum: gram-positive and gram-negative bacteria IGGALGNALNGLGTWANMMNGGGFVNQWQVYANKGKINQYRPY (SEQ ID NO: 16) Unclassified Colicin V Escherichia coli Active against Escherichia coli (as well as closely related bacteria), Enterobacteriaceae MRTLTLNELDSVSGGASGRDIAMAIGTLSGQFVAGGIGAAAGGVAGGAIYDYASTHKPNPAMSPSGLGGTIKQKPEGIPSEAWNYAAGRLCNWSPNNLSDVCL (SEQ ID NO: 17)

In some cases, the bacteriocin is colicin, pyocin, or microcin produced by gram-negative bacteria. In some cases, the bacteriocin is colicin. The colicin may be a group A colicin (eg, uses the Tol system to penetrate the outer membrane of the target bacterium) or a group B colicin (eg, uses the Ton system to penetrate the outer membrane of the target bacterium). In some cases, the bacteriocin is microcin. The microcin may be a class I microcin (eg, <5 kDa, has post-translational modifications) or a class II microcin (eg, 5-10 kDa, with or without post-translational modifications). In some cases, class II microcin is class IIa microcin (eg, requires more than one gene for the synthesis and assembly of functional peptides) or class IIb microcin (eg, linear peptides with or without post-translational modifications at the C-terminus). In some cases, the bacteriocin is pyocin. In some cases, the pyocin is R-pyocin, F-pyocin, or S-pyocin.

In some cases, the bacteriocin is a class I, class II, class III, or class IV bacteriocin produced by gram-positive bacteria. In some cases, the modulating agent includes a class I bacteriocin (eg, lanthionine-containing antibiotics (lantibiotics) produced by Gram-positive bacteria). Class I bacteriocins or lantibiotics may be a low molecular weight peptide (eg, less than about 5 kDa) and may have amino acid residues with post-translational modifications (eg, lanthionine, β-methyllanthionine, or dehydrogenated amino acids).

In some cases, the bacteriocin is a class II bacteriocin (eg, non-lantibiotic compounds produced by gram-positive bacteria). Many of them are positively charged lanthionine-free peptides that, unlike lantibiotics, do not undergo extensive post-translational modification. Class II bacteriocin may belong to one of the following subclasses: “pediocin-like” bacteriocins (eg, pediocin PA-1 and carnobacteriocin X (class IIa)); two-peptide bacteriocins (eg, lactacin F and ABP-118 (class IIb)); circular bacteriocins (eg, carnocyclin A and enterocin AS-48 (class IIc)) or unmodified linear non-pediocin-like bacteriocins (eg, epidermycin NI01 and lactococcin A (class IId)).

In some cases, the bacteriocin is a class III bacteriocin (eg, produced by gram-positive bacteria). Class III bacteriocins can have a molecular weight greater than 10 kDa and may be thermo-unstable proteins. Class III bacteriocins can be further subdivided into group IIIA and group IIIB bacteriocins. Group IIIA bacteriocins include bacteriolytic enzymes that kill susceptible strains through cell wall lysis, such as enterolysin A. Group IIIB bacteriocins include nonlytic proteins, such as caseicin 80, gelveticin J, and lactacin B.

In some cases, the bacteriocin is a class IV bacteriocin (eg, produced by gram-positive bacteria). Class IV bacteriocins are a group of complex proteins associated with other lipid or carbohydrate moieties that appear to be required for activity. They are relatively hydrophobic and thermostable. Examples of class IV bacteriocins are leuconocin S, lactocin 27 and lactocin S.

In some cases, the bacteriocin is an R-type bacteriocin. R-type bacteriocins are contractile bactericidal protein complexes. Some R-type bacteriocins have a contractile structure similar to a phage appendage. The C-terminal region of the phage filament protein determines the specificity of target binding. They can attach to target cells via a protein that binds to a receptor, such as filament filament. Attachment is followed by contraction of the outer sheath and insertion of the core through the membrane of the target bacterium. Penetration of the core leads to rapid depolarization of the cell membrane potential and instantaneous cell death. Contact with one R-type bacteriocin particle can lead to cell death. An R-type bacteriocin, for example, may be heat labile, moderately acid fast, trypsin resistant, centrifugable, detectable by electron microscopy, or a combination of these properties. Other R-type bacteriocins may be complex molecules comprising numerous proteins, polypeptides or subunits, and may resemble the structure of the appendage of bacteriophages of the family Myoviridae. In naturally occurring R-type bacteriocins, the subunit structures may be encoded by the bacterial genome, such as that of C. difficile or P. aeruginosa , and form R-type bacteriocins that act as a natural defense against other bacteria. In some cases, the R-type bacteriocin is pyocin. In some cases, the pyocin is R-pyocin, F-pyocin, or S-pyocin.

In some cases, the bacteriocin is a functionally active variant of the bacteriocins described herein. In some cases, the bacteriocin variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a specific region or in the entire sequence, the sequence of a bacteriocin described herein, or a naturally occurring bacteriocin.

In some cases, a bacteriocin can be bioengineered according to standard methods to modulate its biological activity, eg, increase or decrease, or regulate, or to determine its target microorganisms. In other cases, the bacteriocin is produced by the translational apparatus (for example, ribosome, etc.) of the microbial cell. In some cases, bacteriocin is synthesized chemically. Some bacteriocins can be formed from a polypeptide precursor. The precursor polypeptide may be cleaved (eg, processed by a protease) to form the bacteriocin polypeptide itself. Thus, in some cases the bacteriocin is produced from a precursor polypeptide. In some other cases, the bacteriocin includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The bacteriocins described herein can be formulated for any of the routes of use described herein. The compositions disclosed herein may contain any number or type (e.g., classes) of bacteriocins, such as at least about any number of 1 bacteriocin, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 100 or more bacteriocins. Suitable concentrations of each bacteriocin in the compositions described herein depend on factors such as the effectiveness, stability of the bacteriocin, the amount of different types of bacteriocin in the compositions, the composition and methods of use of the composition. In some cases, the concentration of each bacteriocin in the liquid composition is from about 0.01 ng/ml to about 100 mg/ml. In some cases, the concentration of each bacteriocin in the solid composition is from about 0.01 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of bacteriocins, the concentration of each type of bacteriocin may be the same or different. In some cases, the bacteriocin is provided in a composition containing a bacterial cell that secretes the bacteriocin. In some cases, the bacteriocin is provided in a composition containing a polypeptide (eg, a polypeptide isolated from a bacterial cell).

Bacteriocins can neutralize (eg, kill) at least one microorganism other than the individual bacterial cell in which the polypeptide is produced, including cells clonally related to the bacterial cell and other microbial cells. Thus, a bacterial cell can exert cytotoxic or growth inhibitory effects on a variety of microorganisms through the secretion of bacteriocins. In some cases, a bacteriocin specifically targets and destroys one or more species of bacteria in the host through the formation of pores in the cytoplasmic membrane, cell wall disruption (eg, peptidoglycanase activity), or nuclease activity (eg, DNase activity, 16S rRNase activity, or tRNase activity).

In some cases, the bacteriocin is characterized by neutralizing activity. The neutralizing activity of bacteriocins may include, but is not limited to, inhibition of microbial proliferation or cytotoxicity. Some bacteriocins are characterized by cytotoxic activity, and thus can destroy microorganisms, such as bacteria, yeasts, algae, etc. Some bacteriocins can inhibit the proliferation of microorganisms, such as bacteria, yeasts, algae, etc., e.g. , through cell cycle arrest.

In some cases, the bacteriocin is characterized by destructive activity. The mechanism of destruction of bacteriocins is specific to each group of bacteriocins. In some cases, the bacteriocin is characterized by narrow-spectrum biological activity. Bacteriocins are known for their very high activity against their target strains. The activity of some bacteriocins is limited to strains that are closely related to the bacteriocin-producing strain (narrow-spectrum biological activity). In some cases, the bacteriocin is characterized by broad-spectrum biological activity against a wide range of genera.

In some cases, bacteriocins interact with a receptor molecule or docking molecule on the cell membrane of the target bacterium. For example, nisin is extremely active against its target bacterial strains, demonstrating antimicrobial activity even at single-digit nanomolar concentrations. The nisin molecule has been shown to bind to lipid II, which is the main transporter of peptidoglycan subunits from the cytoplasm to the cell wall.

In some cases, the bacteriocin is characterized by antifungal activity. A number of bacteriocins with antiyeast or antifungal activity have been identified. For example, bacteriocins from Bacillus have been shown to have neutralizing activity against certain yeast strains (see, for example, Adetunji and Olaoye, Malaysian Journal of Microbiology 9:130-13, 2013). In another example, a peptide from Enterococcus faecalis has been shown to have neutralizing activity against Candida species (see, for example, Shekh and Roy, BMC Microbiology 12:132, 2012). In another example, bacteriocins from Pseudomonas have been shown to have neutralizing activity against fungi such as Curvularia lunata , Fusarium spp., Helminthosporium spp., and Biopolaris spp. (see, for example, Shalani and Srivastava, The Internet Journal of Microbiology Volume 5 Number 2, 2008). In another example, botricidin AJ1316 and alirin B1 from B. subtilis were shown to have antifungal activity.

A modulating agent comprising a bacteriocin described herein may be brought into contact with the target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) concentration of the bacteriocin within target host; (b) achieving a target level (eg, a predetermined or threshold level) of bacteriocin concentration within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of bacteriocin concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/and (e) modulating the fitness of the target host.

(b) Lysines

The modulating agent described herein may include lysine (eg, also known as murein hydrolase or peptidoglycan autolysin). Any lysine suitable for inhibiting the bacterium present in the host may be used. In some cases, the lysine is lysine that can be produced naturally by the bacterial cell. In some cases, the lysine is lysine that can be produced naturally by a bacteriophage. In some cases, lysine is obtained from a phage that inhibits a bacterium living in the host. In some cases, lysine is constructed from naturally occurring lysine. In some cases, the lysine is engineered for secretion by the host bacterium, for example, by introducing a signal peptide into the lysine. In some cases, lysine is used in combination with phage choline. In some cases, lysine is expressed by a recombinant bacterial host that is not susceptible to lysine. In some cases, lysine is used to inhibit gram-positive or gram-negative bacteria living in the host.

The lysine may be of any class of lysine and may have one or more substrate specificities. For example, lysine may be a glycosidase, endopeptidase, carbopeptidase, or a combination thereof. In some cases, lysine cleaves the β-1-4 glycosidic bond in a sugar moiety of the cell wall, the amide bond connecting sugar and peptide moieties of the bacterial cell wall, and/or peptide bonds between peptide moieties of the cell wall. Lysine may belong to one or more specific lysine groups depending on the cleavage site in the peptidoglycan. In some cases, lysine is N -acetyl-β-D-muramidase (e.g., lysozyme), lytic transglycosylase, N -acetyl-β-D-glucosaminidase, N -acetylmuramyl-L-alanine amidase, L,D-endopeptidase, D, D-endopeptidase, D,D-carboxypeptidase, L,D-carboxypeptidase or L,D-transpeptidase. Non-limiting examples of lysines and their activity forms are given in Table 5.

Table 5. Examples of lysines

Target bacterium Producer Lysines Activity Subsequence S. pneumoniae Cp1 Cpl-1 Muramidase MVKKNDLFVDVSSHNGYDITGILEQMGTTNTIIKISESTTYLNPCLSAQVEQSNPIGFYHFARFGGDVAEAEREAQFFLDNVPMQVKYLVLDYEDDPSGDAQANTNACLRFMQMIADAGYKPIYYSYKPFTHDNVDYQQILAQFPNSLWIAGYGLNDGTANFEYFPSMDGIRWWQYSSNPFDKNIVLLDDEEDDKP KTAGTWKQDSKGWWFRRNNGSFPYNKWEKIGGVWYYFDSKGYCLTSEWLKDNEKWYYLKDNGAMATGWVLVGSEWYYMDDSGAMVTGWVKYKNNWYYMTNERGNMVSNEFIKSGKGWYFMNTNGELADNPSFTKEPDGLITVA (SEQ ID NO: 18) S. pneumoniae Dp-1 Pal Amidase MGVDIEKGVAWMQARKGRVSYSMDFRDGPDSYDCSSSMYYALRSAGASSAGWAVNTEYMHAWLIENGYELISENAPWDAKRGDIFIWGRKGASAGAGGHTGMFIDSDNIIHCNYAYDGISVNDHDERWYYAGQPYYYVYRLTNANAQPAEKKLGWQKDATGFWYARANGTYPKDEFEYIEENKSWFYFDDQ GYMLAEKWLKHTDGNWYWFDRDGYMATSWKRIGESWYYFNRDGSMVTGWIKYYDNWYYCDATNGDMKSNAFIRYNDGWYLLLPDGRLADKPQFTVEPDGLITAKV (SEQ ID NO: 19) S. pyogenes C1 C1 Amidase But. B. anthracis γ PlyG Amidase MEIQKKLVDPSKYGTKCPYTMKPKYITVHNTYNDAPAENEVSYMISNNNEVSFHIAVDDKKAIQGIPLERNAWACGDGNGSGNRQSISVEICYSKSGGDRYYKAEDNAVDVVRQLMSMYNIPIENVRTHQSWSGKYCPHRMLAEGRWGAFIQKVKNGNVATTSPTKQNIIQSGAFSPYETPDVMGALTSLKMTADFI LQSDGLTYFISKPTSDAQLKAMKEYLDRKGWWYEVK (SEQ ID NO: 20) B. anthracis Ames prophage PlyPH Amidase But. E. faecalis and E. faecium Phi1 PlyV12 Amidase But. S. aureus MR11 MV-L Endopeptidase and amidase MQAKLTKKEFIEWLKTSEGKQFNVDLWYGFQCFDYANAGWKVLFGLLLKGLGAKDIPFANNFDGLATVYQNTPDFLAQPGDMVVFGSNYGAGYGHVAWVIEATLDYIIVYEQNWLGGGWTDRIEQPGWGWEKVTRRQHAYDFPMWFIRPNFKSETAPRSIQSPTQASKKETAKPQPKAVEL KIIKDVVKGYDLPKRGGNPKGIVIHNDAGSKGATAEAYRNGLVNAPLSRLEAGIAHSYVSGNTVWQALDESQVGWHTANQLGNKYYYGIEVCQSMGADNATFLKNEQATFQECARLLKKWGLPANRNTIRLHNEFTSTSCPHRSSVLHTGFDPVTRGLLPEDKQLQLKDYFIKQIRVYMDGKIPVATVSN ESSASSNTVKPVASAWKRNKYGTYYMEENARFTNGNQPITVRKIGPFLSCPVAYQFQPGGYCDYTEVMLQDGHVWVGYTWEGQRYYLPIRTWNGSAPPNQILGDLWGEIS (SEQ ID NO: 21) S. pyogenes C1 PlyC Amidase But. S. agalactiae B30 Lysine GBS Muramidase and endopeptidase MVINIEQAIAWMASRKGKVTYSMDYRNGPSSYDCSSSVYFALRSAGASDNGWAVNTEYEHDWLIKNGYVLIAENTNWNAQRGDIFIWGKRGASAGAFGHTGMFVDPDNIIHCNYGYNSITVNNHDEIWGYNGQPYVYAYRYSGKQSNAKVDNKSVVSKFEKELDVNTPLSNSNMPYYEATISEDYYVESKPD VNSTDKELLVAGTRVRVYEKVKGWARIGAPQSNQWVEDAYLIDATDM (SEQ ID NO: 22) S. aureus P68 Lys16 Endopeptidase But. S. aureus K LysK Amidase and endopeptidase MAKTQAEINKRLDAYAKGTVDSPYRVKKATSYDPSFGVMEAGAIDADGYYHAQCQDLITDYVLWLTDNKVRTWGNAKDQIKQSYGTGFKIHENKPSTVPKKGWIAVFTSGSYEQWGHIGIVYDGGNTSTFTILEQNWNGYANKKPTKRVDNYYGLTHFIEIPVKAGTTVKKETAKKS ASKTPAPKKKATLKVSKNHINYTMDKRGKKPEGMVIHNDAGRSSGQQYENSLANAGYARYANGIAHYYGSEGYVWEAIDAKNQIAWHTGDGTGANSGNFRFAGIEVCQSMSASDAQFLKNEQAVFQFTAEKFKEWGLTPNRKTVRLHMEFVPTACPHRSMVLHTGFNPVTQGRPSQAIMNKLKDYFIKQIK NYMDKGTSSSTVVKDGKTSSASTPATRPVTGSWKKNQYGTWYKPENATFVNGNQPIVTRIGSPFLNAPVGGNLPAGATIVYDEVCIQAGHIWIGYNAYNGNRVYCPVRTCQGVPPNQIPGVAWGVFK (SEQ ID NO: 23) L. monocytogenes A118 Ply118 Amidase MTSYYYSRSLANVNKLADNTKAAARKLLDWSESNGIEVLIYETIRTKEQQAANVNSGASQTMRSYHLVGQALDFVMAKGKTVDWGAYRSDKGKKFVAKAKSLGFEWGGDWSGFVDNPHLQFNYKGYGTDTFGKGASTSNSSKPSADTNTNSLGLVDYMNLNKLDSSFANRKKLATSYGIKNYSGTAT QNTTLLAKLKAGKPHTPASKNTYYTENPRKVKTLVQCDLYKSVDFTTKNQTGGTFPPGTVFTISGMGKTKGGTPRLKTKSGYYLTANTKFVKKI (SEQ ID NO: 24) L. monocytogenes A511 Ply511 Amidase MVKYTVENKIIAGLPKGKLKGANFVIAHETANSKSTIDNEVSYMTRNWKNAFVTHFVGGGGRVVQVANVNYVSWGAGQYANSYSYAQVELCRTSNATTFKKDYEVYCQLLVDLAKKAGIPITLDSGSKTSDKGIKSHKWVADKLGGTTHQDPYAYLSSWGISKAQFASDLAKVSGGGNTGTAPAKPSTPAPK PSTPSTNLDKLGLVDYMNAKKMDSSYSNRDKLAKQYGIANYSGTASQNTTLLSKIKGGAPKPSTPAPKPSTSTAKKIYFPPNKGNWSVYPTNKAPVKANAIGAINPTKFGGLTYTIQKDRGNGVYEIQTDQFGRVQVYGAPSTGAVIKK (SEQ ID NO: 25) L. monocytogenes A500 Ply500 Endopeptidase MALTEAWLIEKANRKLNAGGMYKITSDKTRNVIKKMAKEGIYLCVAQGYRSTAEQNALYAQGRTKPGAIVTNAKGGQSNHNYGVAVDLCLYTNDGKDVIWESTTSRWKKVVAAMKAEGFKWGGDWKSFKDYPHFELCDAVSGEKIPAATQNTNTNSNRYEGKVIDSAPLLPKMDFKSSPFRMYKVGTE FLVYDHNQYWYKTYIDDKLYYMYKSFCDVVAKKDAKGRIKVRIKSAKDLRIPVWNNIKLNSGKIKWYAPNVKLAWYNYRRGYLELWYPNDGWYYTAEYFLK (SEQ ID NO: 26) S. pneumoniae ΦDp-1 Pal, S Endopeptidase and amidase But. S. agalactiae Prophage LambdaSa1 LambdaSa1 Glycosidase MVINIEQAIAWMASRKGKVTYSMDYRNGPSSYDCSSSVYFALRSAGASDNGWAVNTEYEHDWLIKNGYVLIAENTNWNAQRGDIFIWGKRGASAGAFGHTGMFVDPDNIIHCNYGYNSITVNNHDEIWGYNGQPYVYAYRYARKQSNAKVDNQSVVSKFEKELDVNTPLSNSNMPYYEATISEDYYVESKPD VNSTDKELLVAGTRVRVYEKVKGWARIGAPQSNQWVEDAYLIDATDM (SEQ ID NO: 27) S. agalactiae Prophage LambdaSa2 LambdaSa2 Glycosidase and endopeptidase MEINTEIAIAWMSARQGKVSYSMDYRDGPNSYDCSSSVYYALRSAGASSAGWAVNTEYMHDWLIKNGYELIAENVDWNAVRGDIAIWGMRGHSSGAGGHVVMFIDPENIIHCNWANNGITVNNYNQTAAASGWMYCYVYRLKSGASTQGKSLDTLVKETLAGNYGNGEARKAVLGNQYEAVMSVINGKTTTNQKTVDQLV QEVIAGKHGNGEARKKSLGSQYDAVQKRVTELLKKQPSEPFKAQEVNKPTETKTSQTELTGQATATKEEGDLSFNGTILKKAVLDKILGNCKKHDILPSYALTILHYEGLWGTSAVGKADNNWGGMTWTGQGNRPSGVTVTQGSARPSNEGGHYMHYASVDDFLTDWFYLLRAGGSYKVSGAKTFSEAIK GMFKVGGAVYDYAASGFDSYIVGASSRLKAIEAENGSLDKFDKATDIGDGSKDKIDITIEGIEVTINGITYELTKKPV (SEQ ID NO: 28) S. uberis Prophage (ATCC700407) Ply700 Amidase MTDSIQEMRKLQSIPVRYDMGDRYGNDADRDGRIEMDCSSAVSKALGISMTNNTETLQQALPAIGYGKIHDAVDGTFDMQAYDVIIWAPRDGSSSLGAFGHVLIATSPTTAIHCNYGSDGITENDYNYIWEINGRPREIVFRKGVTQTQATVTSQFERELDVNARLTVSDKPYYEATLSEDYYVEAGPRIDSQDKEL IKAGTRVRVYEKLNGWSRINHPESAQWVEDSYLVDATEM (SEQ ID NO: 29) S. suis SMP LySMP Glycosidase and endopeptidase But. B. anthracis Bcp1 PlyB Muramidase But. S. aureus Phi11 and Phi12 Lysine Phi11 Amidase and endopeptidase MQAKLTKNEFIEWLKTSEGKQFNVDLWYGFQCFDYANAGWKVLFGLLLKGLGAKDIPFANNFDGLATVYQNTPDFLAQPGDMVVFGSNYGAGYGHVAWVIEATLDYIIVYEQNWLGGGWTDGIEQPGWGWEKVTRRQHAYDFPMWFIRPNFKSETAPRSVQSPTQAPKKETAKPQPKAVEL KIIKDVVKGYDLPKRGSNPKGIVIHNDAGSKGATAEAYRNGLVNAPLSRLEAGIAHSYVSGNTVWQALDESQVGWHTANQIGNKYYYGIEVCQSMGADNATFLKNEQATFQECARLLKKWGLPANRNTIRLHNEFTSTSCPHRSSVLHTGFDPVTRGLLPEDKRLQLKDYFIKQIRAYMDGKIPVATVSNESSASSN TVKPVASAWKRNKYGTYYMEESARFTNGNQPITVRKVGPFLSCPVGYQFQPGGYCDYTEVMLQDGHVWVGYTWEGQRYYLPIRTWNGSAPPNQILGDLWGEIS (SEQ ID NO: 30) S. aureus ΦH5 LysH5 Amidase and endopeptidase MQAKLTKKEFIEWLKTSEGKQYNADGWYGFQCFDYANAGWKALFGLLLKGVGAKDIPFANNFDGLATVYQNTPDFLAQPGDMVVFGSNYGAGYGHVAWVIEATLDYIIVYEQNWLGGGWTDGVQQPGSGWEKVTRRQHAYDFPMWFIRPNFKSETAPRSVQSPTQASKKETAKPQPK AVELKIIKDVVKGYDLPKRGSNPNFIVIHNDAGSKGATAEAYRNGLVNAPLSRLEAGIAHSYVSGNTVWQALDESQVGWHTANQIGNKYGYGIEVCQSMGADNATFLKNEQATFQECARLLKKWGLPANRNTIRLHNEFTSTSCPHRSSVLHTGFDPVTRGLLPEDKRLQLKDYFIKQIRAYMDGKIPVATVSNDSS ASSNTVKPVASAWKRNKYGTYYMEESARFTNGNQPITVRKVGPFLSCPVGYQFQPGGYCDYTEVMLQDGHVWVGYTWEGQRYYLPIRTWNGSAPPNQILGDLWGEIS (SEQ ID NO: 31) S. warneri ΦWMY LysWMY Amidase and endopeptidase MKTKAQAKSWINSKIGKGIDWDGMYGYQCMDEAVDYIHHVTDGKVTMWGNAIDAPKNNFQGLCTVYTNTPEFRPAYGDVIVWSYGTFATYGHIAIVVNPDPYGDLQYITVLEQNWNGNGIYKTEFATIRTHDYTGVSHFIRPKFADEVKETAKTVNKLSVQKKIVTPKNSVERIKNYVKTSGY INGEHYELYNRGHKPKGVVIHNTAGTASATQEGQRLTNMTFQQLANGVAHVYIDKNTIYETLPEDRIAWHVAQQYGNTEFYGIEVCGSRNTDKEQFLANEQVAFQEAARRLKSWGMRANRNTVRLHHTFSSTECPDMSMLLHTGYSMKNGKPTQDITNKCADYFMKQINAYIDGKQPTSTVVGSSSSNKLKAK NKDKSTGWNTNEYGTLWKKEHATFTCGVRQGIVTRTTGPFTSCPQAGVLYYGQSVNYDTVCKQDGYVWISWTTSDGYDVWMPIRTWDRSTDKVSEIWGTIS (SEQ ID NO: 32) Streptococci (GBS) ΦNCTC 11261 PlyGBS Muramidase and endopeptidase MATYQEYKSRSNGNAYDIDGSFGAQCWDGYADYCKYLGLPYANCTNTGYARDIWEQRHENGILNYFDEVEVMQAGDVAIFMVVDGVTPYSHVAIFDSDAGGYGWFLGQNQGGANGAYNIVKIPYSATYPTAFRPKVFKNAVTVTGNIGLNKGDYFIDVSAYQQADLTTTCQQAGTTKTIIKVSESI AWLSDRHQQQANTSDPIGYYHFGRFGGDSALAQREADLFLSNLPSKKVSYLVIDYEDSASADKQANTNAVIAFMDKIASAGYKPIYYSYKPFTLNNIDYQKIIAKYPNSIWIAGYPDYEVRTEPLWEFFPSMDGVRWWQFTSVGVAGGLDKNIVLLADDSSKMDIPKVDKPQELTFYQKLATNTKLDNSNVP YYEATLSTDYYVESKPNASSADKEFIKAGTRVRVYEKVNGWSRINHPESAQWVEDSYLVNATDM (SEQ ID NO: 33) C. perfringens Φ3626 Ply3626 Amidase But. C. difficile ΦCD27 Lysine CD27 Amidase But. E. faecalis Φ1 PlyV12 Amidase But. A. naeslundii ΦAv-1- Lysine Av-1 Putative amidase/muramidase But. L. gasseri ΦgaY LysgaY Muramidase But. S. aureus ΦSA4 LysSA4 Amidase and endopeptidase But. S. haemolyticus ΦSH2 SH2 Amidase and endopeptidase But. B. thuringiensis ΦBtCS33 PlyBt33 Amidase But. L. monocytogenes ΦP40 PlyP40 Amidase But. L. monocytogenes ΦFWLLm3 LysZ5 Amidase MVKYTVENKIIAGLPKGKLKGANFVIAHETANSKSTIDNEVSYMTRNWQNAFVTHFVGGGGRVVQVANVNYVSWGAGQYANSYSYAQVELCRTSNATTFKKDYEVYCQLLVDLAKKAGIPITLDSGSKTSDKGIKSHKWVADKLGGTTHQDPYAYLSSWGISKAQFASDLAKVSGGGNTGTAPAKPSTPSTN (SEQ ID NO: 34) B. cereus ΦBPS13 LysBPS13 Amidase MAKREKYIFDVEAEVGKAAKSIKSLEAELSKLQKLNKEIDATGGDRTEKEMLATLKAAKEVNAEYQKMQRILKDLSKYSGKVSRKEFNDSKVINNAKTSVQGGKVTDSFGQMLKNMERQINSVNKQFDNHRKAMVDRGQQYTPHLKTNRKDSQGNSNPSMMGRNKSTTQDMEKAVDKFLNGQNEATT GLNQALYQLKEISKLNRRSESLSRRASASGYMSFQQYSNFTGDRRTVQQTYGGLKTANRERVLELSGQATGISKELDRLNSKKGLTAREGEERKKLMRQLEGIDAELTARKKLNSSLDETTSNMEKFNQSLVDAQVSVKPERGTMRGMMYERAPAIALAIGGAITATIGKLYSEGGNHSKAMRPDEMYVGQQ TGAVGANWRPNRTATMRSGLGNHLGFTGQEMMEFQSNYLSANGYHGAEDMKAATTGQATFARATGLGSDEVKDFFNTAYRSGGIDGNQTKQFQNAFLGAMKQSGAVGREKDQLKALNGILSSMSQNRTVSNQDMMRTVGLQSAISSSGVASLQGTKGGALMEQLDNGIREGFNDPQMRVLFGQGTKYQGMGGRAAL RKQMEKGISDPDNLNTLIDASKASAGQDPAEQAEVLATLASKMGVNMSSDQARGLIDLQPSGKLTKENIDKVMKEGLKEGSIESAKRDKAYSESKASIDNSSEAATAKQATELNDMGSKLRQANAALGGLPAPLYTAIAAVVAFTAAVAGSALMFKGASWLKGGMASKYGGKGGKGGKGGGTGGGGGAGGAAATGAGAAAGAGG VGAAAAGEVGAGVAAGGAAAGAAAGGSKLAGVGKGFMKGAGKLMLPLGILMGASEIMQAPEEAKGSAIGSAVGGIGGGIAGGAATGAIAGSFLGPIGTAVGGIAGGIAGGFAGSSLGETIGGWFDSGPKEDASAADKAKADASAAALAAAAGTSGAVGSSALQSQMAQGITGAPNMSQVGSMASALGISSGAMASALGISSGQENQIQ TMTDKENTNTKKANEAKKGDNLSYERENISMYERVLTRAEQILAQARAQNGIMGVGGGGTAGAGGGINGFTGGGKLQFLAAGQKWSSSNLQQHDLGFTDQNLTAEDLDKWIDSKAPQGSMMRGMGATFLKAGQEYGLDPRYLIAHAAEESGWGTSKIARDKGNFFGIGAFDDSPYSSAYEFKDGTGSAAERGIMGGAKWISE KYYGKGNTTLDKMKAAGYATNASWAPNIASIMAGAPTGSGSGNVTATINVNVKGDEKVSDKLKNSSDMKKAGKDIGSLLGFYSREMTIA (SEQ ID NO: 35) S. aureus ΦGH15 LysGH15 Amidase and endopeptidase MAKTQAEINKRLDAYAKGTVDSPYRIKKATSYDPSFGVMEAGAIDADGYYHAQCQDLITDYVLWLTDNKVRTWGNAKDQIKQSYGTGFKIHENKPSTVPKKGWIAVFTSGSYQQWGHIGIVYDGGNTSTFTILEQNWNGYANKKPTKRVDNYYGLTHFIEIPVKAGTTVKKETAKKSAS KTPAPKKKATLKVSKNHINYTMDKRGKKPEGMVIHNDAGRSSGQQYENSLANAGYARYANGIAHYYGSEGYVWEAIDAKNQIAWHTGDGTGANSGNFRFAGIEVCQSMSASDAQFLKNEQAVFQFTAEKFKEWGLTPNRKTVRLHMEFVPTACPHRSMVLHTGFNPVTQGRPSQAIMNKLKDYFIKQIKNY MDKGTSSSTVVKDGKTSSASTPATRPVTGSWKKNQYGTWYKPENATFVNGNQPIVTRIGSPFLNAPVGGNLPAGATIVYDEVCIQAGHIWIGYNAYNGDRVYCPVRTCQGVPPNHIPGVAWGVFK (SEQ ID NO: 36) S. aureus ΦvB SauS-PLA88 HydH5 Endopeptidase and glycosidase But. E. faecalis ΦF168/08 Lys168 Endopeptidase But. E. faecalis ΦF170/08 Lys170 Amidase But. S. aureus ΦP-27/HP P-27/HP Undefined But. C. perfringens ΦSM101 Psm Muramidase But. C. sporogenes Φ8074-B1 CS74L Amidase MKIGIDMGHTLSGADYGVVGLRPESVLTREVGTKVIYKLQKLGHVVVNCTVDKASSVSESLYTRYYRANQANVDLFISIHFNATPGGTGTEVYTYAGRQLGEATRIRQEFKSLGLRDRGTKDGSGLAVIRNTKAKAMLVECCFCDNPNDMKLYNSESFSNAIVKGITGKLPNGESGNNNQGGNKVKAVVIYNEGADR RGAEYLADYLNCPTISNSRTFDYSCVEHVYAVGGKKEQYTKYLKTLLSGANRYDTMQQILNFINGGK (SEQ ID NO: 37) S. typhimurium ΦSPN1S SPN1S Glycosidase MDINQFRRASGINEQLAARWFPHITTAMNEFGITKPDDQAMFIAQVGHESGGFTRLQENFNYSVNGLSGFIRAGRITPDQANALGRKTYEKSLPLERQRAIANLVYSKRMGNNGPGDGWNYRGRGLIQITGLNNYRDCGNGLKVDLVAQPELLAQDEYAAARSAAWFFSSKGCMKYTGDLVRVTQIINGGQNGIDDRRTRY AAARKVLAL (SEQ ID NO: 38) C. michiganensis ΦCMP1 CMP1 Peptidase But. C. michiganensis ΦCN77 CN77 Peptidase MGYWGYPNGQIPNDKMALYRGCLLRADAAAQAYALQDAYTRATGKPLVILEGYRDLTRQKYLRNLYLSGRGNIAAVPGLSNHGWGLACDFAAPLNSSGSEEHRWMRQNAPLFGFDWARGKADNEPWHWEYGNVPVSRWASLDVTPIDRNDMADITEGQMQRIAVILLDTEIQTPLGPRLVKHALGDALLLGQANANSIAEVPDK TWDVLVDHPLAKNEDGTPLKVRLGDVAKYEPLEHQNTRDAIAKLGTLQFTDKQLATIGAGVKPIDEASLVKKIVDGVRALFGRAAA (SEQ ID NO: 39) A. baumannii ΦAB2 LysAB2 Glycosidase MILTKDGFSIIRNELFGGKLDQTQVDAINFIVAKATESGLTYPEAAYLLATIYHETGLPSGYRTMQPIKEAGSDSYLRSKKYYPYIGYGYVQLTWKENYERIGKLIGVDLIKNPEKALEPLIAIQIAIKGMLNGWFTGVGFRRKRPVSKYNKQQYVAARNIINGKDKAELIAKYAIIFERALRSL (SE Q ID NO: 40) B. cereus ΦB4 LysB4 Endopeptidase MAMALQTLIDKANRKLNVSGMRKDVADRTRAVITQMHAQGIYICVAQGFRSFAEQNALYAQGRTKPGSIVTNARGGQSNHNYGVAVDLCLYTQDGSDVIWTVEGNFRKVIAAMKAQGFKWGGDWVSFKDYPHFELYDVVGGQKPPADNGGAVDNGGGSGSTGGSGGGSTGGGSTGGGYDSSWFTKETGTFVTNTSI KLRTAPFTSADVIATLPAGSPVNYNGFGIEYDGYVWIRQPRSNGYGYLATGESKGGKRQNYWGTFK (SEQ ID NO: 41) P. aeruginosa ΦKMV KMV45 Undefined But. C. tyrobutyricum ΦCTP1 Ctp1l Glycosidase MKKIADISNLNGNVDVKLLFNLGYIGIIAKASEGGTFVDKYYKQNYTNTKAQGKITGAYHFANFSTIAKAQQEANFFLNCIAGTTPDFVVLDLEQQCTGDITDACLAFLNIVAKKFKCVVYCNSSFIKEHLNSKICAYPLWIANYGVATPAFTLWTKYAMWQFTEKGQVSGISGYIDFSYITDEFIKYI KGEDEVENLVVYNDGADQRAAEYLADRLACPTINNARKFDYSNVKNVYAVGGNKEQYTSYLTTLIAGSTRYTTMQAVLDYIKNLK (SEQ ID NO: 42) P. aeruginosa ΦEL EL188 Transglycosylase But. P. aeruginosa ΦKZ KZ144 Transglycosylase But. S. aureus Staphylococcal virus 187 Ply187 Cell wall hydrolase MALPKTGKPTAKQVVDWAINLIGSGVDVDGYYGRQCWDLPNYIFNRYWNFKTPGNARDMAWYRYPEGFKVFRNTSDFVPKPGDIAVWTGGNYNWNTWGHTGIVVGPSTKSYFYSVDQNWNNNSSYVGSPAAKIKHSYFGVTHFVRPAYKAEPKPTPPAQNNPAPKDPEPSKKPESNKPIYKVVTK ILFTTAHIEHVKANRFVHYITKSDNHNNKPNKIVIKNTNTALSTIDVYRYRDELDKDEIPHFFVDRLNVWACRPIEDSINGYHDSVVLSITETRTALSDNFKMNEIECLSLAESILKANNKKMSASNIIVDNKAWRTFKLHTGKDSLKSSSFTSKDYQKAVNELIKLFNDKDKLLNNKPKDVVERIRIRTIVKENTKFVP SELKPRNNIRDKQDSKIDRVINNYTLKQALNIQYKLNPKPQTSNGVSWYNASVNQIKSAMDTTKIFNNNVQVYQFLKLNQYQGIPVDKLNKLLVGKGTLANQGHAFADGCKKYNINEIYLIAHRFLESANGTSFFASGKTGVYNYFGIGAFDNNPNNAMAFARSHGWTSPTKAIIGGAEFVGKGY FNVGQNTLYRMRWNPQKPGTHQYATDISWAKVQAQMISAMYKEIGLTGDYFIYDQYKK (SEQ ID NO: 43) P. fluorescens ΦOBP OBPgp279 Glycosidase But. L. monocytogenes ΦP35 PlyP35 Amidase MARKFTKAELVAKAEKKVGGLKPDVKKAVLSAVKEAYDRYGIGIIVSQGYRSIAEQNGLYAQGRTKPGNIVTNAKGGQSNHNFGVAVDFAIDLIDDGKIDSWQPSATIVNMMKRRGFKWGGDWKSFTDLPHFEACDWYRGERKYKVDTSEWKKKENINIVIKDVGYFQDKPQFLNSKSVRQWK HGTKVKLTKHNSHWYTGVVKDGNKSVRGYIYHSMAKVTSKNSDGSVNATINAHAFCWDNKKLNGGDFINLKRGFKGITHPASDGFYPLYFASRKKTFYIPRYMFDIKK (SEQ ID NO: 44) L. fermentum ΦPYB5 Lyb5 Muramidase But. S. pneumoniae ΦCP-7 Cpl-7 Muramidase MVKKNDLFVDVASHQGYDISGILEEAGTTNTIIKVSESTSYLNPCLSAQVSQSNPIGFYHFAWFGGNEEEAEARYFLDNVPTQVKYLVLDYEDHASASVQRNTTACLRFMQIIAEAGYTPIYYSYKPFTLDNVDYQQILAQFPNSLWIAGYGLNDGTANFEYFPSMDGIRWWQYSSNPFDKNIVLLDDE KEDNINNENTLKSLTTVANEVIQGLWGNGQERYDSLANAGYDPQAVQDKVNEILNAREIADLTTVANEVIQGLWGNGQERYDSLANAGYDPQAVQDKVNEILNAREIADLTTVANEVIQGLWGNGQERYDSLANAGYDPQAVQDKVNELLS (SEQ ID NO: 45) P. chlororaphis 201 Φ2-1 201ϕ2-1gp229 Glycosidase But. S. enterica ΦPVP-SE1) PVP-SE1gp146 Glycosidase But. Corynebacterium ΦBFK20 BKF20 Amidase But. E. faecalis EFAP-1 EFAL-1 Amidase MKLKGILLSVVTTFGLLFGATNVQAYEVNNEFNLQPWEGSQQLAYPNKIILHETANPRATGRNEATYMKNNWFNAHTTAIVGDGGIVYKVAPEGNVSWGAGNANPYAPVQIELQHTNDPELFKANYKAYVDYTRDMGKKFGIPMTLDQGGSLWEKGVVSHQWVTDFVWGDHTDPYGYLAKMGISKAQ LAHDLANGVSGNTATPTPKPDKPKPTQPSKPSNKKRFNYRVDGLEYVNGMWQIYNEHLGKIDFNWTENGIPVEVVDKVNPATGQPTKDQVLKVGDYFNFQENSTGVVQEQTPYMGYTLSHVQLPNEFIWLFTDSKQALMYQ (SEQ ID NO: 46) Lactobacilli LambdaSA2 LysA, LysA2, and Lysga Y Undefined But. S. aureus SAL-1 Undefined But.

In some cases, the lysine is a functionally active variant of the lysines described herein. In some cases, the lysine variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a specific region or entire sequence, the sequence of a lysine described herein, or a naturally occurring lysine.

In some cases, lysine can be bioengineered to modulate its biological activity, eg, increase or decrease, or regulate, or to determine its target microorganism. In some cases, lysine is produced by the translational apparatus (for example, ribosome, etc.) of the microbial cell. In some cases, lysine is synthesized chemically. In some cases, lysine is formed from a polypeptide precursor. The precursor polypeptide may be cleaved (eg, processed by a protease) to form the lysine polypeptide itself. Thus, in some cases, lysine is produced from a precursor polypeptide. In some cases, lysine includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The lysines described herein may be formulated for any of the routes of use described herein. The compositions disclosed herein may contain any number or type (e.g., classes) of lysines, such as at least about any number of 1, 2, 3, 4, 5, 10, 15, 20, or more lysines . The appropriate concentration of each lysine in the composition depends on factors such as the potency, stability of the lysine, the number of different lysines, the composition and methods of use of the composition. In some cases, the concentration of each lysine in the liquid composition is from about 0.1 ng/ml to about 100 mg/ml. In some cases, the concentration of each lysine in the solid composition is from about 0.1 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of lysines, the concentration of each type of lysines may be the same or different.

A modulatory agent comprising lysine described herein may be brought into contact with the target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of lysine concentration within target host; (b) achieving a target level (eg, a predetermined or threshold level) of lysine concentration within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of lysine concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/and (e) modulating the fitness of the target host.

(c) Antimicrobial peptides

The modulatory agent described herein may include an antimicrobial peptide (AMP). Any AMP suitable for inhibiting a microorganism living in the host can be used. AMPs are a diverse group of molecules that are divided into subgroups based on their amino acid composition and structure. AMPs can be derived from or produced in any organism that naturally produces AMPs, including AMPs derived from plants (e.g., copsin), insects (e.g., drozocin, scorpion peptide (e.g., Uy192, UyCT3, D3, D10, Uy17, Uy192), mastoparan, poneratoxin, cecropin, moricin, mellitin), frogs (e.g. magainin, dermaseptin, aurein) and mammals (e.g. cathelicidins, defensins and protegrins). For example, the AMP may be a scorpion peptide such as Uy192 (5'-FLSTIWNGIKGLL-3'; SEQ ID NO: 193), UyCT3 (5'-LSAIWSGIKSLF-3'; SEQ ID NO: 194), D3 (5'- LWGKLWEGVKSLI-3'; SEQ ID NO: 195) and D10 (5'-FPFLKLSLKIPKSAIKSAIKRL-3'; SEQ ID NO: 196), Uy17 (5'-ILSAIWSGIKGLL-3'; SEQ ID NO: 197). Other non-limiting examples of AMPs are shown in Table 6.

Table 6. Examples of antimicrobial peptides

Type Characteristic Illustrative AMP Subsequence Anionic peptides Rich in glutamic and aspartic acid Dermsidin SSLLEKGLDGAKKAVGGLGKLGKDAVEDLESVGKGAVHDVKDVLDSVL (SEQ ID NO: 47) Linear cationic α-helical peptides Do not have cysteine Cecropin A KWKLFKKIEKVGQNIRDGIIKAGPAVAVVGQATQIAK (SEQ ID NO: 48) Andropin MKYFSVLVVLTLILAIVDQSDAFINLLDKVEDALHTGAQAGFKLIRPVERGATPKKSEKPEK (SEQ ID NO: 49) Moricin MNILKFFFVFIVAMSLVSCSTAAPAKIPIKAIKTVGKAVGKGLRAINIASTANDVFNFLKPKKRKH (SEQ ID NO: 50) Ceratotoxin MANLKAVFLICIVAFIALQCVVAEPAAEDSVVVKRSIGSALKKALPVAKKIGKIALPIAKAALPVAAGLVG (SEQ ID NO: 51) Cationic peptides enriched with specific amino acids Rich in proline, arginine, phenylalanine, glycine, tryptophan Abetzin MKVVIFIFALLATICAAFAYVPLPNVPQPGRRFPTFPGQGPFNPKIKWPQGY (SEQ ID NO: 52) Apidecines KNFALAILVVTFVVAVFGNTNLDPPTRPTRLRREAKPEAEPGNNRPVYIPQPRPPHPRLRREAEPEAEPGNNRPVYIPQPRPPHPRLRREAELEAEPGNNRPVYISQPRPPHPRLRREAEPEAEPGNNRPVYIPQPRPPHPRLRREAELEAEPGNNRPVYISQPRPPHPRLRREAEPEAEPGNNRPVYIPQPRPPHPRLRREAEPEAEPG NNRPVYIPQPRPPHPRLRREAEPEAEPGNNRPVYIPQPRPPHPRLRREAKPEAKPGNNRPVYIPQPRPPHPRI (SEQ ID NO: 53) Profenin METQRASLCLGRWSLWLLLLALVVPSASAQALSYREAVLRAVDRLNEQSSEANLYRLLELDQPPKADEDPGTPKPVSFTVKETVCPRPTRRPPELCDFKENGRVKQCVGTVTLDQIKDPLDITCNEGVRRFPWWWPFLRRPRLRRQAFPPPPNVPGPRFPPPNVPGPRFPPPNFPGPRFPPPNFPGPRFPPPNFPGPPFPPPIFPGPWFPPPPP FRPPPFGPPRFPGRR (SEQ ID NO: 54) Indolicidin MQTQRASLSLGRWSLWLLLLGLVVPSASAQALSYREAVLRAVDQLNELSSEANLYRLLELDPPPKDNEDLGTRKPVSFTVKETVCPRTIQQPAEQCDFKEKGRVKQCVGTVTLDPSNDQFDLNCNELQSVILPWKWPWWPWRRG (SEQ ID NO: 55) Anionic and cationic peptides that contain cysteine and form disulfide bonds Contain 1-3 disulfide bonds Protegrin METQRASLCLGRWSLWLLLLALVVPSASAQALSYREAVLRAVDRLNEQSSEANLYRLLELDQPPKADEDPGTPKPVSFTVKETVCPRPTRQPPELCDFKENGRVKQCVGTVTLDQIKDPLDITCNEVQGVRGGRLCYCRRRFCVCVGRG (SEQ ID NO: 56) Tachyplesins KWCFRVCYRGICYRRCR (SEQ ID NO: 57) Defensins MKCATIVCTIAVVLAATLLNGSVQAAPQEEAALSGGANLNTLLDELPEETHHAALENYRAKRATCDLASGFGVGSSLCAAHCIARRYRGGYCNSKAVCVCRN (SEQ ID NO: 58) Drozomycin MMQIKYLFALFAVLMLVVLGANEADADCLSGRYKGPCAVWDNETCRRVCKEEGRSSGHCSPSLKCWCEGC (SEQ ID NO: 59)

AMP can be active against any number of target microorganisms. In some cases, AMP may exhibit forms of antibacterial and/or antifungal activity. In some cases, an AMP may have narrow-spectrum biological activity or broad-spectrum biological activity. For example, some AMPs specifically target and kill only a few species of bacteria or fungi, while others are active against both Gram-negative and Gram-positive bacteria as well as fungi.

In addition, AMP may function through a number of known mechanisms of action. For example, the cytoplasmic membrane is a common target of AMPs, but AMPs can also interfere with DNA and protein synthesis, protein folding, and cell wall synthesis. In some cases, AMPs with a net cationic charge and amphipathic nature disrupt bacterial membranes, leading to cell lysis. In some cases, AMPs can enter cells and interact with intracellular targets to disrupt DNA, RNA, protein, or cell wall synthesis. In addition to killing microorganisms, AMPs have demonstrated a number of immunomodulatory functions that are involved in clearing infection, including the ability to alter host gene expression, act as chemokines and/or induce chemokine production, inhibit lipopolysaccharide-induced proinflammatory cytokine production, promote wound healing, and modulate responses. dendritic cells and cells involved in the adaptive immune response.

In some cases, the AMP is a functionally active variant of the AMPs described herein. In some cases, the AMP option is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a specific region or in the entire sequence, the sequence of an AMP described herein, or a naturally occurring AMP.

In some cases, an AMP can be bioengineered to modulate its biological activity, for example, to increase or decrease or regulate, or to determine its target microorganism. In some cases, AMP is produced by the translational machinery (e.g., ribosome, etc.) of the cell. In some cases, AMPs are synthesized chemically. In some cases, AMP is formed from a polypeptide precursor. The precursor polypeptide may be cleaved (eg, processed by a protease) to form the AMP polypeptide itself. Thus, in some cases, AMP is produced from a precursor polypeptide. In some cases, the AMP includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The AMPs described herein may be formulated for any of the routes of administration described herein. The compositions disclosed herein may contain any number or type (e.g., classes) of AMPs, such as at least about any number of 1 AMP, 2, 3, 4, 5, 10, 15, 20, or more AMPs . The appropriate concentration of each AMP in the composition depends on factors such as potency, stability of the AMP, the number of different AMPs in the composition, composition, and methods of use of the composition. In some cases, the concentration of each AMP in the liquid composition is from about 0.1 ng/ml to about 100 mg/ml. In some cases, the concentration of each AMP in the solid composition is from about 0.1 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of AMP, the concentration of each type of AMP may be the same or different.

A modulatory agent comprising an AMP described herein may be brought into contact with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of the concentration of the AMP within target host; (b) achieving a target level (eg, a predetermined or threshold level) of AMP concentration within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of AMP concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/or (e) modulating the fitness of the target host.

(d) C-rich nodule peptides

The modulating agent described herein may include C-rich nodule peptide (NCR peptide). NCR peptides are produced in certain species of leguminous plants and play an important role in the mutualistic nitrogen-fixing symbiosis of plants with bacteria from the family Rhizobiaceae (nodule bacteria), which leads to the formation of root nodules in which plant cells contain thousands of intracellular endosymbionts. NCR peptides have antimicrobial properties by controlling the irreversible process of bacterial terminal differentiation, such as permeabilization of the bacterial membrane, disruption of cell division, or inhibition of protein synthesis. For example, nodule cells of Medicago truncatula infected with Sinorhizobium meliloti produce hundreds of NCR peptides that drive the irreversible process of bacterial differentiation into large polyploid nitrogen-fixing bacteroids. Non-limiting examples of NCR peptides are shown in Table 7.

Table 7. Examples of NCR peptides

Name Peptide sequence Producer >gi|152218086|gb|ABS31477,1| NCR 340 MTKIVVFIYVVILLLTIFHVSAKKKRYIECETHEDCSQVFMPPFVMRCVIHECKIFNGEHLRY (SEQ ID NO: 60) Medicago truncatula >gi|152218084|gb|ABS31476,1| NCR 339 MAKIMKFVYNMIPFLSIFIITLQVNVVVCEIDADCPQICMPPYEVRCVNHRCGWVNTDDDSLFLTQEFTRSKQYIIS (SEQ ID NO: 61) Medicago truncatula >gi|152218082|gb|ABS31475,1| NCR 338 MYKVVESIFIRYMHRKPNMTKFFKFVYTMFILISLFLVVTNANAHNCTDISDCCSSNHCSYEGVSLCMNGQCICIYE (SEQ ID NO: 62) Medicago truncatula >gi|152218080|gb|ABS31474,1| NCR 337 MVETLRLFYIMILFVSLCLVVVDGESKLEQTCSEDFECYIKNPHVPFGHLRCFEGFCQQLNGPA (SEQ ID NO: 63) Medicago truncatula >gi|152218078|gb|ABS31473,1| NCR 336 MAKIVNFVYSMIVFLFLFLVATKAARGYLCVTDSHCPPHMCPPGMEPRCVRRMCKCLPIGWRKYFVP (SEQ ID NO: 64) Medicago truncatula >gi|152218076|gb|ABS31472,1| NCR 335 MQIGKNMVETPKLDYVIIFFFLYFFFRQMIILRLNTTFRPLNFKMLRFWGQNRNIMKHRGQKVHFSLILSDCKTNKDCPKLRRANVRCRKSYCVPI (SEQ ID NO: 65) Medicago truncatula >gi|152218074|gb|ABS31471,1| NCR 334 MLRLYLVSYFLLKRTLLVSYFSYFSTYIIECKTDNDCPISQLKIYAWKCVKNGCHLFDVIPMYE (SEQ ID NO: 66) Medicago truncatula >gi|152218072|gb|ABS31470,1| NCR 333 MAEILKFVYIVILFVSLLLIVVASERECVTDDDCEKLYPTNEYRMMCDSGYCMNLLNGKIIYLLCLKKKKFLIIISVLL (SEQ ID NO: 67) Medicago truncatula >gi|152218070|gb|ABS31469,1| NCR 332 MAEIIKFVYIMILCVSLLLIEVAGEECVTDADCDKLYPDIRKPLMCSIGECYSLYKGKFLSSIISKTSFSLMVYNVVTLVICLRLAYISLLLKFL (SEQ ID NO: 68) Medicago truncatula >gi|152218068|gb|ABS31468,1| NCR 331 MAEILKDFYAMNLFIFLIILPAKIRGETLSLTHPKCHHIMLPSLFITEVFQRVTDDGCPKPVNHLRVVKCIEHICEYGYNYRPDFASQIPESTKMPRKRE (SEQ ID NO: 69) Medicago truncatula >gi|152218066|gb|ABS31467,1| NCR 330 MVEILKNFYAMNLFIFLIILAVKIRGAHFPCVTDDDCPKPVNKLRVIKCIDHICQYARNLPDFASEISESTKMPCKGE (SEQ ID NO: 70) Medicago truncatula >gi|152218064|gb|ABS31466,1| NCR 329 MFHAQAENMAKVSNFVCIMILFLALFFITMNDAARFECREDSHCVTRIKCVLPRKPECRNYACGCYDSNKYR (SEQ ID NO: 71) Medicago truncatula >gi|152218062|gb|ABS31465,1| NCR 328 MQMRQNMATILNFVFVIILFISLLLVVTKGYREPFSSFTEGPTCKEDIDCPSISCVNPQVPKCIMFECHCKYIPTTLK (SEQ ID NO: 72) Medicago truncatula >gi|152218060|gb|ABS31464,1| NCR 327 MATILMYVYITILFISILTVLTEGLYEPLYNFRRDPCRRNIDCPSYLCVAPKVPRCIMFECHCKDIPSDH (SEQ ID NO: 73) Medicago truncatula >gi|152218058|gb|ABS31463,1| NCR 326 MTTSLKFVYVAILFLSLLLVVMGGIRRFECRQDSDCPSYFCEKLTVPKCFWSKCYCK (SEQ ID NO: 74) Medicago truncatula >gi|152218056|gb|ABS31462,1| NCR 325 MTTSLKFVYVAILFLSLLLVVMGGIRKKECRQDSDCPSYFCEKLTIAKCIHSTCLCK (SEQ ID NO: 75) Medicago truncatula >gi|152218054|gb|ABS31461,1| NCR 324 MQIGKNMVETPKLVYFIILFLSIFLCITVSNSSFSQIFNSACKTDKDCPKFGRVNVRCRKGNCVPI (SEQ ID NO: 76) Medicago truncatula >gi|152218046|gb|ABS31457,1| NCR 320 MTAILKKFINAVFLFIVLFLATTNVEDFVGGSNDECVYPDVFQCINNICKCVSHHRT (SEQ ID NO: 77) Medicago truncatula >gi|152218044|gb|ABS31456,1| NCR 319 MQKRKNMAQIIFYVYALIILFSPFLAARLVFVNPEKPCVTDADCDRYRHESAIYSDMFCKDGYCFIDYHHDPYP (SEQ ID NO: 78) Medicago truncatula >gi|152218042|gb|ABS31455,1| NCR 318 MQMRKNMAQILFYVYALLILFTTPFLVARIMVVNPNNPCVTDADCQRYRHKLATRMICNQGFCLMDFTHDPYAPSLP (SEQ ID NO: 79) Medicago truncatula >gi|152218040|gb|ABS31454,1| NCR 317 MNHISKFVYALIIFLSIYLVVLDGLPISCKDHFECRRKINILRCIYRQEKPMCINSICCTCVKLL (SEQ ID NO: 80) Medicago truncatula >gi|152218038|gb|ABS31453,1| NCR 316 MQREKNMAKIFEFVYAMIIFILLFLVEKNVVAYLKFECKTDDDCQKSLLKTYVWKCVKNECYFFAKK (SEQ ID NO: 81) Medicago truncatula >gi|152218036|gb|ABS31452,1| NCR 315 MAGIIKFVHVLIIFLSLFHVVKNDDGSFCFKDSDCPDEMCPSPLKEMCYFLQCKCGVDTIA (SEQ ID NO: 82) Medicago truncatula >gi|152218034|gb|ABS31451,1| NCR 314 MANTHKLVSMILFIFLFLASNNVEGYVNCETDADCPPSTRVKRFKCVKGECRWTRMSYA (SEQ ID NO: 83) Medicago truncatula >gi|152218032|gb|ABS31450,1| NCR 313 MQRRKKKAQVVMFVHDLIICIYLFIVITTRKTDIRCRFYYDCPRLEYHFCECIEDFCAYIRLN (SEQ ID NO: 84) Medicago truncatula >gi|152218030|gb|ABS31449,1| NCR 312 MAKVYMFVYALIIFVSPFLLATFRTRLPCEKDDDCPEAFLPPVMKCVNRFCQYEILE (SEQ ID NO: 85) Medicago truncatula >gi|152218028|gb|ABS31448,1| NCR 310 MIKQFSVCYIQMRRNMTTILKFPYIMVICLLLHVAAYEDFEKEIFDCKKDGDCDHMCVTPGIPKCTGYVCFCFENL(SEQ ID NO: 86) Medicago truncatula >gi|152218026|gb|ABS31447,1| NCR 309 MQRSRNMTTIFKFAYIMIICVFLLNIAAQEIENGIHPCKKNEDCNHMCVMPGLPWCHENNLCFCYENAYGNTR (SEQ ID NO: 87) Medicago truncatula >gi|152218024|gb|ABS31446,1| NCR 304 MTIIIKFVNVLIIFLSLFHVAKNDDNKLLLSFIEEEGFLCFKDSDCPYNMCPSPLKEMCYFIKCVCGVYGPIRERRLYQSHNPMIQ (SEQ ID NO: 88) Medicago truncatula >gi|152218022|gb|ABS31445,1| NCR 303 MRKNMTKILMIGYALMIFIFLSIAVSITGNLARASRKKPVDVIPCIYDHDCPRKLYFLERCVGRVCKYL (SEQ ID NO: 89) Medicago truncatula >gi|152218020|gb|ABS31444,1| NCR 301 MAHKLVYAITLFIFLFLIANNIEDDIFCITDNDCPPNTLVQRYRCINGKCNLSFVSYG (SEQ ID NO: 90) Medicago truncatula >gi|152218018|gb|ABS31443,1| NCR 300 MDETLKFVYILILFVSLCLVVADGVKNINRECTQTSDCYKKYPFIPWGKVRCVKGRCRLDM (SEQ ID NO: 91) Medicago truncatula >gi|152218016|gb|ABS31442,1| NCR 290 MAKIIKFVYVLAIFFSLFLVAKNVNGWTCVEDSDCPANICQPPMQRMCFYGECACVRSKFCT (SEQ ID NO: 92) Medicago truncatula >gi|152218014|gb|ABS31441,1| NCR 289 MVKIIKFVYFMTLFLSMLLVTTKEDGSVECIANIDCPQIFMLPFVMRCINFRCQIVNSEDT (SEQ ID NO: 93) Medicago truncatula >gi|152218012|gb|ABS31440,1| NCR 286 MDEILKFVYTLIIFFSLFFAANNVDANIMNCQSTFDCPRDMCSHIRDVICIFKKCKCAGGRYMPQVP (SEQ ID NO: 94) Medicago truncatula >gi|152218008|gb|ABS31438,1| NCR 278 MQRRKNMANNHMLIYAMIICLFPYLVVTFKTAITCDCNEDCLNFFTPLDNLKCIDNVCEVFM (SEQ ID NO: 95) Medicago truncatula >gi|152218006|gb|ABS31437,1| NCR 266 MVNILKFIYVIIFFILMFFVLIDVDGHVLVECIENRDCEKGMCKFPFIVRCLMDQCKCVRIHNLI (SEQ ID NO: 96) Medicago truncatula >gi|152218004|gb|ABS31436,1| NCR 265 MIIQFSIYYMQRRKLNMVEILKFSHALIIFFLLSALVTNANIFFCSTDEDCTWNLCRQPWVQKCRLHMCSCEKN (SEQ ID NO: 97) Medicago truncatula >gi|152218002|gb|ABS31435,1| NCR 263 MDEVFKFVYVMIIFPFLILDVATNAEKIRRCFNDAHCPPDMCTLGVIPKCSRFTICIC (SEQ ID NO: 98) Medicago truncatula >gi|152218000|gb|ABS31434,1| NCR 244 MHRKPNMTKFFKFVYTMFILISLFLVVTNANANNCTDTSDCSSNHCSYEGVSLCMNGQCICIYE (SEQ ID NO: 99) Medicago truncatula >gi|152217998|gb|ABS31433,1| NCR 239 MQMKKMATILKFVYLIILLIYPLLVVTEESHYMKFSICKDDTDCPTLFCVLPNVPKCIGSKCHCKLMVN (SEQ ID NO: 100) Medicago truncatula >gi|152217996|gb|ABS31432,1| NCR 237 MVETLRLFYIMILFVSLYLVVVDGVSKLAQSCSEDFECYIKNPHAPFGQLRCFEGYCQRLDKPT (SEQ ID NO: 101) Medicago truncatula >gi|152217994|gb|ABS31431,1| NCR 228 MTTFLKVAYIMIICVFVLHLAAQVDSQKRLHGCKEDRDCDNICSVHAVTKCIGNMCRCLANVK (SEQ ID NO: 102) Medicago truncatula >gi|152217992|gb|ABS31430,1| NCR 224 MRINRTPAIFKFVYTIIIYLFLLRVVAKDLPFNICEKDEDCLEFCAHDKVAKCMLNICFCF (SEQ ID NO: 103) Medicago truncatula >gi|152217990|gb|ABS31429,1| NCR 221 MAEILKILYVFIIFLSLILAVISQHPFTPCETNADCKCRNHKRPDCLWHKCYCY (SEQ ID NO: 104) Medicago truncatula >gi|152217988|gb|ABS31428,1| NCR 217 MRKSMATILKFVYVIMLFIYSLFVIESFGHRFLIYNNCKNDTECPNDCGPHEQAKCILYACYCVE (SEQ ID NO: 105) Medicago truncatula >gi|152217986|gb|ABS31427,1| NCR 209 MNTILKFIFVVFLFLSIFLSAGNSKSYGPCTTLQDCETHNWFEVCSCIDFECKCWSLL (SEQ ID NO: 106) Medicago truncatula >gi|152217984|gb|ABS31426,1| NCR 206 MAEIIKFVYIMILCVSLLLIAEASGKECVTDADCENLYPGNKKPMFCNNTGYCMSLYKEPSRYM (SEQ ID NO: 107) Medicago truncatula >gi|152217982|gb|ABS31425,1| NCR 201 MAKIIKFVYIMILCVSLLLIVEAGGKECVTDVDCEKIYPGNKKPLICSTGYCYSLYEEPPRYHK (SEQ ID NO: 108) Medicago truncatula >gi|152217980|gb|ABS31424,1| NCR 200 MAKVTKFGYIIIHFLSLFFLAMNVAGGRECHANSHCVGKITCVLPQKPECWNYACVCYDSNKYR (SEQ ID NO: 109) Medicago truncatula >gi|152217978|gb|ABS31423,1| NCR 192 MAKIFNYVYALIMFLSLFLMGTSGMKNGCKHTGHCPRKMCGAKTTKCRNNKCQCV (SEQ ID NO: 110) Medicago truncatula >gi|152217976|gb|ABS31422,1| NCR 189 MTEILKFVCVMIIFISSFIVSKSLNGGGKDKCFRDSDCPKHMCPSSLVAKCINRLCRCRRPELQVQLNP (SEQ ID NO: 111) Medicago truncatula >gi|152217974|gb|ABS31421,1| NCR 187 MAHIIMFVYALIYALIIFSSLFVRDGIPCLSDDECPEMSHYSFKCNNKICEYDLGEMSDDDYYLEMSRE (SEQ ID NO: 112) Medicago truncatula >gi|152217972|gb|ABS31420,1| NCR 181 MYREKNMAKTLKFVYVIVLFLSLFLAAKNIDGRVSYNSFIALPVCQTAADCPEGTRGRTYKCINNKCRYPKLLKPIQ (SEQ ID NO: 113) Medicago truncatula >gi|152217970|gb|ABS31419,1| NCR 176 MAHIFNYVYALLVFLSLFLMVTNGIHIGCDKDRDCPKQMCHLNQTPKCLKNICKCV (SEQ ID NO: 114) Medicago truncatula >gi|152217968|gb|ABS31418,1| NCR 175 MAEILKCFYTMNLFIFLIILPAKIREHIQCVIDDDCPKSLNKLLIIKCINHVCQYVGNLPDFASQIPKSTKMPYKGE (SEQ ID NO: 115) Medicago truncatula >gi|152217966|gb|ABS31417,1| NCR 173 MAYISRIFYVLIIFLSLFFVVINGVKSLLLIKVRSFIPCQRSDDCPRNLCVDQIIPTCVWAKCKCKNYND (SEQ ID NO: 116) Medicago truncatula >gi|152217964|gb|ABS31416,1| NCR 172 MANVTKFVYIAIYFLSLFFIAKNDATATFCHDDSHCVTKIKCVLPRTPQCRNEACGCYHSNKFR (SEQ ID NO: 117) Medicago truncatula >gi|152217962|gb|ABS31415,1| NCR 171 MGEIMKFVYVMIIYLFMFNVATGSEFIFTKKLTSCDSSKDCRSFLCYSPKFPVCKRGICECI (SEQ ID NO: 118) Medicago truncatula >gi|152217960|gb|ABS31414,1| NCR 169 MGEMFKFIYTFILFVHLFLVVIFEDIGHIKYCGIVDDCYKSKKPLFKIWKCVENVCVLWYK (SEQ ID NO: 119) Medicago truncatula >gi|152217958|gb|ABS31413,1| NCR 165 MARTLKFVYSMILFLSLFLVANGLKIFCIDVADCPKDLYPLLYKCIYNKCIVFTRIPPFFDWI (SEQ ID NO: 120) Medicago truncatula >gi|152217956|gb|ABS31412,1| NCR 159 MANITKFVYIAILFLSLFFIGMNDAAILECREDSHCVTKIKCVLPRKPECRNNACTCYKGGFSFHH (SEQ ID NO: 121) Medicago truncatula >gi|152217954|gb|ABS31411,1| NCR 147 MQRVKKMSETLKFVYVLILFISIFHVVIVCDSIYFPVSRPCITDKDCPNMKHYKAKCRKGFCISSRVR (SEQ ID NO: 122) Medicago truncatula >gi|152217952|gb|ABS31410,1| NCR 146 MQIRKIMSGVLKFVYAIILFLFLFLVAREVGGLETIECETDGDCPRSMIKMWNKNYRHKCIDGKCEWIKKLP (SEQ ID NO: 123) Medicago truncatula >gi|152217950|gb|ABS31409,1| NCR 145 MFVYDLILFISLILVVTGINAEADTSCHSFDDCPWVAHHYRECIEGLCAYRILY (SEQ ID NO: 124) Medicago truncatula >gi|152217948|gb|ABS31408,1| NCR 144 MQRRKKSMAKMLKFFFAIILLLSLFLVATEVGGAYIECEVDDDCPKPMKNSHPDTYYKCVKHRCQWAWK (SEQ ID NO: 125) Medicago truncatula >gi|152217946|gb|ABS31407,1| NCR 140 MFVYTLIIFLFPSHVITNKIAIYCVSDDDCLKTFTPLDLKCVDNVCEFNLRCKGKCGERDEKFVFLKALKKMDQKLVLEEQGNAREVKIPKKLLFDRIQVPTPATKDQVEEDDYDDDDEEEEEEEDDVDMWFHLPDVVCH (SEQ ID NO: 126) Medicago truncatula >gi|152217944|gb|ABS31406,1| NCR 138 MAKFSMFVYALINFLSLFLVETAITNIRCVSDDDCPKVIKPLVMKCIGNYCYFFMIYEGP (SEQ ID NO: 127) Medicago truncatula >gi|152217942|gb|ABS31405,1| NCR 136 MAHKFVYAIILFIFLFLVAKNVKGYVVCRTVDDCPPDTRDLRYRCLNGKCKSYRLSYG (SEQ ID NO: 128) Medicago truncatula >gi|152217940|gb|ABS31404,1| NCR 129 MQRKKNMGQILIFVFALINFLSPILVEMTTTTIPCTFIDDCPKMPLVVKCIDNFCNYFEIK (SEQ ID NO: 129) Medicago truncatula >gi|152217938|gb|ABS31403,1| NCR 128 MAQTLMLVYALIIFTSLFLVVISRQTDIPCKSDDACPRVSSHHIECVKGFCTYWKLD (SEQ ID NO: 130) Medicago truncatula >gi|152217936|gb|ABS31402,1| NCR 127 MLRRKNTVQILMFVSALLIYIFLFLVITSSANIPCNSDSDCPWKIYYTYRCNDGFCVYKSIDPSTIPQYMTDLIFPR (SEQ ID NO: 131) Medicago truncatula >gi|152217934|gb|ABS31401,1| NCR 122 MAVILKFVYIMIIFLFLLYVVNGTRCNRDEDCPFICTGPQIPKCVSHICFCLSSGKEAY (SEQ ID NO: 132) Medicago truncatula >gi|152217932|gb|ABS31400,1| NCR 121 MDAILKFIYAMFLFLFLFVTTRNVEALFECNRDFVCGNDDECVYPYAVQCIHRYCKCLKSRN (SEQ ID NO: 133) Medicago truncatula >gi|152217930|gb|ABS31399.1| NCR 119 MQIGRKKMGETPKLVYVIILFLSIFLCTNSSFSQMINFRGCKRDKDCPQFRGVNIRCRSGFCTPIDS (SEQ ID NO: 134) Medicago truncatula >gi|152217928|gb|ABS31398,1| NCR 118 MQMRKNMAQILFYVYALLILFSPFLVARIMVVNPNNPCVTDADCQRYRHKLATRMVCNIGFCLMDFTHDPYAPSLP (SEQ ID NO: 135) Medicago truncatula >gi|152217926|gb|ABS31397,1| NCR 111 MYVYYIQMGKNMAQRFMFIYALIIFLSQFFVVINTSDIPNNSNRNSPKEDVFCNSNDDCPTILYYVSKCVYNFCEYW (SEQ ID NO: 136) Medicago truncatula >gi|152217924|gb|ABS31396,1| NCR 103 MAKIVNFVYSMIIFVSLFLVATKGGSKPFLTPYPCNTGSDCPQNMCPPGYKPGCEDGYCNHCYKRW (SEQ ID NO: 137) Medicago truncatula >gi|152217922|gb|ABS31395,1| NCR 101 MVRTLKFVYVIILILSLFLVAKGGGKKIYCENAASCPRLMYPLVYKCLDNKCVKFMMKSRFV (SEQ ID NO: 138) Medicago truncatula >gi|152217920|gb|ABS31394,1| NCR 96 MARTLKFVYAVILFLSLFLVAKGDDVKIKCVVAANCPDLMYPLVYKCLNGICVQFTLTFPFV (SEQ ID NO: 139) Medicago truncatula >gi|152217918|gb|ABS31393,1| NCR 94 MSNTLMFVITFIVLVTLFLGPKNVYAFQPCVTTADCMKTLKTDENIWYECINDFCIPFPIPKGRK (SEQ ID NO: 140) Medicago truncatula >gi|152217916|gb|ABS31392,1| NCR 93 MKRVVNMAKIVKYVYVIIIFLSLFLVATKIEGYYYKCFKDSDCVKLLCRIPLRPKCMYRHICKCKVVLTQNNYVLT (SEQ ID NO: 141) Medicago truncatula >gi|152217914|gb|ABS31391,1| NCR 90 MKRGKNMSKILKFIYATLVLYLFLVVTKASDDECKIDGDCPISWQKFHTYKCINQKCKWVLRFHEY (SEQ ID NO: 142) Medicago truncatula >gi|152217912|gb|ABS31390,1| NCR 88 MAKTLNFMFALILFISLFLVSKNVAIDIFVCQTDADCPKSELSMYTWKCIDNECNLFKVMQQMV (SEQ ID NO: 143) Medicago truncatula >gi|152217910|gb|ABS31389,1| NCR 86 MANTHKLVSMILFIFLFLVANNVEGYVNCETDADCPPSTRVKRFKCVKGECRWTRMSYA (SEQ ID NO: 144) Medicago truncatula >gi|152217908|gb|ABS31388,1| NCR 77 MAHFLMFVYALITCLSLFLVEMGHLSIHCVSVDDCPKVEKPITMKCINNYCKYFVDHKL (SEQ ID NO: 145) Medicago truncatula >gi|152217906|gb|ABS31387,1| NCR 76 MNQIPMFGYTLIIFFSLFPVITNGDRIPCVTNGDCPVMRLPLYMRCITYSCELFFDGPNLCAVERI (SEQ ID NO: 146) Medicago truncatula >gi|152217904|gb|ABS31386,1| NCR 74 MRKDMARISLFVYALIIFFSLFFVLTNGELEIRCVSDADCPLFPLPLHNRCIDDVCHLFTS (SEQ ID NO: 147) Medicago truncatula >gi|152217902|gb|ABS31385,1| NCR 68 MAQILMFVYFLIIFLSLFLVESIKIFTEHRCRTDADCPARELPEYLKCQGGMCRLLIKKD (SEQ ID NO: 148) Medicago truncatula >gi|152217900|gb|ABS31384,1| NCR 65 MARVISLFYALIIFLFLFLVATNGDLSPCLRSGDCSKDECPSHLVPKCIGLTCYCI (SEQ ID NO: 149) Medicago truncatula >gi|152217898|gb|ABS31383,1| NCR 62 MQRRKNMAQILLFAYVFIISISLFLVVTNGVKIPCVKDTDCPTLPCPLYSKCVDGFCKMLSI (SEQ ID NO: 150) Medicago truncatula >gi|152217896|gb|ABS31382,1| NCR 57 MNHISKFVYALIIFLSVYLVVLDGRPVSCKDHYDCRRKVKIVGCIFPQEKPMCINSMCTCIREIVP (SEQ ID NO: 151) Medicago truncatula >gi|152217894|gb|ABS31381,1| NCR 56 MKSQNHAKFISFYKNDLFKIFQNNDSHFKVFFALIIFLYTYLHVTNGVFVSCNSHIHCRVNNHKIGCNIPEQYLLCVNLFCLWLDY (SEQ ID NO: 152) Medicago truncatula >gi|152217892|gb|ABS31380,1| NCR 54 MTYISKVVYALIIFLSIYVGVNDCMLVTCEDHFDCRQNVQQVGCSFREIPQCINSICKCMKG (SEQ ID NO: 153) Medicago truncatula >gi|152217890|gb|ABS31379,1| NCR 53 MTHISKFVFALIIFLSIYVGVNDCKRIPCKDNNDCNNNWQLLACRFEREVPRCINSICKCMPM (SEQ ID NO: 154) Medicago truncatula >gi|152217888|gb|ABS31378,1| NCR 43 MVQTPKLVYVIVLLLSIFLGMTICNSSFSHFFEGACKSDKDCPKLHRSNVRCRKGQCVQI (SEQ ID NO: 155) Medicago truncatula >gi|152217886|gb|ABS31377,1| NCR 28 MTKILMLLFYAMIVFHSIFLVASYTDECSTDADCEYILCLFPIIKRCIHNHCKCVPMGSIEPMSTIPNGVHKFHIINN (SEQ ID NO: 156) Medicago truncatula >gi|152217884|gb|ABS31376,1| NCR 26 MAKTLNFVCAMILFISLFLVSKNVALYIIECKTDADCPISKLNMYNWRCIKSSCHLYKVIQFMV (SEQ ID NO: 157) Medicago truncatula >gi|152217882|gb|ABS31375,1| NCR 24 MQKEKNMAKTFEFVYAMIIFILLFLVENNFAAYIIECQTDDDCPKSQLEMFAWKCVKNGCHLFGMYEDDDDP (SEQ ID NO: 158) Medicago truncatula >gi|152217880|gb|ABS31374,1| NCR 21 MAATRKFIYVLSHFLFLFLVTKITDARVCKSDKDCKDIIIYRYILKCRNGECVKIKI (SEQ ID NO: 159) Medicago truncatula >gi|152217878|gb|ABS31373,1| NCR 20 MQRLDNMAKNVKFIYVIILLLFIFLVIIVCDSAFVPNSGPCTTDKDCKQVKGYIARCRKGYCMQSVKRTWSSYSR (SEQ ID NO: 160) Medicago truncatula >gi|152217876|gb|ABS31372,1| NCR 19 MKFIYIMILFLSLFLVQFLTCKGLTVPCENPTTCPEDFCTPPMITRCINFICLCDGPEYAEPEYDGPEPEYDHKGDFLSVKPKIINENMMMRERHMMKEIEV (SEQ ID NO: 161) Medicago truncatula >gi|152217874|gb|ABS31371,1| NCR 12 MAQFLMFIYVLIIFLYLFYVEAAMFELTKSTIRCVTDADCPNVVKPLKPKCVDGFCEYT (SEQ ID NO: 162) Medicago truncatula >gi|152217872|gb|ABS31370,1| NCR 10 MKMRIHMAQIIMFFYALIIFLSPFLVDRRSFPSSFVSPKSYTSEIPCKATRDCPYELYYETKCVDSLCTY (SEQ ID NO: 163) Medicago truncatula

Any NCR peptide known in the art is suitable for use in the methods or compositions described herein. Plants that produce NCR peptides include, but are not limited toPisum sativum (peas),Astragalus sinicus (pulse IRLC),Phaseolus vulgaris(beans),Vigna unguiculata (cow peas),Medicago truncatula(alfalfa truncated) AndLotus japonicus. For example, based on the genomic sequenceM. truncatula over 600 potential NCR peptides are predicted, and nearly 150 different NCR peptides have been identified in cells isolated from root nodules using mass spectrometry.

The NCR peptides described herein may be mature or immature NCR peptides. Immature NCR peptides have a C-terminal signal peptide that is required for translocation into the endoplasmic reticulum and is cleaved off after translocation. The N-terminus of the NCR peptide contains a signal peptide that can be cleavable to target the secretory pathway. NCR peptides are typically small peptides with disulfide bridges that stabilize their structures. Mature NCR peptides have a length ranging from about 20 to about 60 amino acids, from about 25 to about 55 amino acids, from about 30 to about 50 amino acids, from about 35 to about 45 amino acids, or any range in between. NCR peptides may contain a conserved sequence of cysteine residues, with the remainder of the peptide sequence being highly variable. NCR peptides typically have approximately four or eight cysteine residues.

NCR peptides can be anionic, neutral or cationic. In some cases, synthetic cationic NCR peptides having a pI greater than about eight have forms of antimicrobial activity. For example, both NCR247 (pI=10.15, RNGCIVDPRCPYQQCRRPLYCRRR; SEQ ID NO: 164) and NCR335 (pI=11.22) are effective against gram-negative and gram-positive bacteria, as well as fungi. In some cases, neutral and/or anionic NCR peptides, such as NCR001 (MAQFLLFVYSLIIFLSLFFGEAAFERTETRMLTIPCTSDDNCPKVISPCHTKCFDGFCGWYIEGSYEGP; SEQ ID NO: 165), do not exhibit forms of antimicrobial activity at pI greater than about 8.

In some cases, NCR peptide is effective in killing bacteria. In some cases, NCR peptide is effective in killing S. meliloti, Xenorhabdus spp, Photorhabdus spp, Candidatus spp, Buchnera spp, Blattabacterium spp, Baumania spp, Wigglesworthia spp, Wolbachia spp, Rickettsia spp, Orientia spp, Sodalis spp, Burkholderia spp, Cupriavidus spp, Frankia spp, Snirhizobium spp, Streptococcus spp, Wolinella spp, Xylella spp, Erwinia spp, Agrobacterium spp, Bacillus spp, Paenibacillus spp, Streptomyces spp, Micrococcus spp, Corynebacterium spp, Acetobacter spp, Cyanobacteria spp, Salmonella spp, Rhodococcus spp, Pseudomonas spp, Lactobacillus spp, Enterococcus spp, Alcaligenes spp, Klebsiella spp, Paenibacillus spp, Arthrobacter spp, Corynebacterium spp, Brevibacterium spp, Thermus spp, Pseudomonas spp, Clostridium spp or Escherichia spp .

In some cases, the NCR peptide is a functionally active variant of the NCR peptide described herein. In some cases, the NCR peptide variant is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical, for example, in a certain region or in the entire sequence, to the sequence of an NCR peptide described herein or a naturally occurring NCR peptide.

In some cases, the NCR peptide can be bioengineered to modulate its biological activity, eg, increase or decrease, or regulate, or to determine its target microorganism. In some cases, the NCR peptide is produced by the translational machinery (e.g., ribosome, etc.) of the cell. In some cases, the NCR peptide is synthesized chemically. In some cases, the NCR peptide is formed from a polypeptide precursor. The polypeptide precursor may be cleaved (eg, processed by a protease) to form the NCR peptide itself. Thus, in some cases, the NCR peptide is produced from a precursor polypeptide. In some cases, an NCR peptide includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The NCR peptide described herein may be formulated for any of the routes of use described herein. The compositions disclosed herein may contain any number or type of NCR peptides, such as at least about any number of 1 NCR peptide, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50 , 100 or more NCR peptides. The appropriate concentration of each NCR peptide in the composition depends on factors such as potency, stability of the NCR peptide, the number of different NCR peptides, composition and methods of use of the composition. In some cases, the concentration of each NCR peptide in the liquid composition is from about 0.1 ng/ml to about 100 mg/ml. In some cases, the concentration of each NCR peptide in the solid composition is from about 0.1 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of NCR peptides, the concentration of each type of NCR peptide may be the same or different.

A modulating agent comprising an NCR peptide described herein can be brought into contact with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) concentration of the peptide NCR within the target host; (b) achieving a target level (eg, a predetermined or threshold level) of concentration of the NCR peptide within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of concentration of the NCR peptide within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/and (e) modulating the fitness of the target host.

(e) Regulatory peptides of bacteriocytes

The modulatory agent described herein may include bacteriocyte regulatory peptide (BRP). BRPs are peptides expressed in insect bacteriocytes. These genes are first expressed at a developmental time point coinciding with the incorporation of symbionts, and their bacteriocyte-specific expression is maintained throughout the life of the insect. In some cases, BRP has a hydrophobic amino-terminal domain that is predicted to be a signal peptide. In addition, some BRPs have a cysteine-rich domain. In some cases, the bacteriocyte regulatory peptide is a bacteriocyte-specific cysteine-rich protein (BCR). Regulatory peptides of bacteriocytes range from approximately 40 to 150 amino acids in length. In some cases, the bacteriocyte regulatory peptide has a length ranging from about 45 to about 145, from about 50 to about 140, from about 55 to about 135, from about 60 to about 130, from about 65 to about 125, from about 70 to about 120, from about 75 to about 115, from about 80 to about 110, from about 85 to about 105, or anywhere in between. Non-limiting examples of BRPs and their activity forms are given in Table 8.

Table 8. Examples of regulatory peptides of bacteriocytes

Name Peptide sequence Family of bacteriocyte-specific cysteine-rich BCR proteins, BCR1 peptide MKLLHGFLIIMLTMHLSIQYAYGGPFLTKYLCDRVCHKLCGDEFVCSCIQYKSLKGLWFPHCPTGKASVVLHNFLTSP (SEQ ID NO: 166) Family of bacteriocyte-specific cysteine-rich BCR proteins, BCR2 peptide MKLLYGFLIIMLTIHLSVQYFESPFETKYNCDTHCNKLCGKIDHCSCIQYHSMEGLWFPHCRTGSAAQMLHDFLSNP (SEQ ID NO: 167) Family of bacteriocyte-specific cysteine-rich BCR proteins, BCR3 peptide MSVRKNVLPTMFVVLLIMSPVTPTSVFISAVCYSGCGSLALVCFVSNGITNGLDYFKSSAPLSTSETSCGEAFDTCTDHCLANFKF (SEQ ID NO: 168) Family of bacteriocyte-specific cysteine-rich BCR proteins, BCR4 peptide MRLLYGFLIIMLTIYLSVQDFDPTEFKGFPPTIEICSKYCAVVCNYTSRPCYCVEAAKERDQWFPYCYD (SEQ ID NO: 169) Family of bacteriocyte-specific cysteine-rich BCR proteins, BCR5 peptide MRLLYGFLIIMLTIHLSVQDIDPNTLRGPYPTKEICSKYCEYNVVCGASLPCICVQDARQLDHWFACCYDGGPEMLM (SEQ ID NO: 170) SP family of secreted proteins, peptide SP1 MKLFVVVVLVAVGIMFVFASDTAAAPTDYEDTNDMISLSSLVGDNSPYVRVSSADSGGSSKTSSKNPILGLLKSVIKLLTKIFGTYSDAAPAMPPIPPALRKNRGMLA (SEQ ID NO: 171) SP family of secreted proteins, SP2 peptide MVACKVILAVAVVFVAAVQGRPGGEPEWAAPIFAELKSVSDNITNLVGLDNAGEYATAAKNNLNAFAESLKTEAAVFSKSFEGKAASASDVFKESTKNFQAVVDTYIKNLPKDLTLKDFTEKSEQALKYMVEHGTEITKKAQGNTETEKEIKEFFKKQIENLIGQGKALQAKIAEAKKA (SEQ ID NO: 172 ) SP family of secreted proteins, SP3 peptide MKTSSSKVFASCVAIVCLASVANALPVQKSVAATTENPIVEKHGCRAHKNLVRQNVVDLKTYDSMLITNEVVQKQSNEVQSSEQSNEGQNSEQSNEGQNSEQSNEVQSSEHSNEGQNSKQSNEGQNSEQSNEVQSSEHSNEGQNSEQSNEVQSSEHSNEGQNSKQSNEGQNSKQSNEVQSSEHWNEGQ NSKQSNEDQNSEQSNEGQNSKQSNEGQNSKQSNEDQNSEQSNEGQNSKQSNEVQSSEQSNEGQNSKQSNEGQSSEQSNEGQNSKQSNEVQSPEEHYDLPDPESSYESEETKGSHESGDDSEHR (SEQ ID NO: 173) SP family of secreted proteins, peptide SP4 MKTIILGLCLFGALFWSTQSMPVGEVAPAVPAVPSEAVPQKQVEAKPETNAASPVSDAKPESDSKPVDAEVKPTVSEVKAESEQKPSGEPKPESDAKPVVASESKPESDPKPAAVVESKPENDAVAPETNNDAKPENAAAPVSENKPATDAKAETELIAQAKPESKPASDLKAEPEAAKPNSEVPVALPLNPTETKATQ QSVETNQVEQAAPAAAQADPAAAPAADPAPAPAAAPVAAEEAKLSESAPSTENKAAEEPSKPAEQQSAKPVEDAVPAASEISETKVSPAVPAVPEVPASPSAPAVADPVSAPEAEKNAEPAKAANSAEPAVQSEAKPAEDIQKSGAVVSAENPKPVEEQKPAEVAKPAEQSKSEAPAEAPKPTEQSAAEEPKKPESANDEK KEQHSVNKRDATKEKKPTDSIMKKQKQKKAN (SEQ ID NO: 174) SP family of secreted proteins, peptide SP5a (SEQ ID NO: 175) SP family of secreted proteins, peptide SP6 MHLFLALGLFIVCGMVDATFYNPRSQTFNQLMERRQRSIPIPYSYGYHYNPIEPSINVLDSLSEGLDSRINTFKPIYQNVKMSTQDVNSVPRTQYQPKNSLYDSEYISAKDIPSLFPEEDSYDYKYLGSPLNKYLTRPSTQESGIAINLVAIKETSVFDYGFPTYKSPYSSDSVWNFGSKIPNTVFEDPQSVESDPNTFKV SSPTIKIVKLLPETPEQESIITTTKNYELNYKTTQETPTEAELYPITSEEFQTEDEWHPMVPKENTTKDESSFITTEEPLTEDKSNSITIEKTQTEDESNSIEFNSIRTEEKSNSITTEENQKEDDESMSTTSQETTTAFNLNDTFDTNRYSSSHESLMLRIRELMKNIADQQNKSQFRTVDNIPAKSQSNLSSDESTNQQFEPQLVNGADTYK (SEQ ID NO: 176) Coleoptericin A, ColA peptide MTRTMLFLACVAALYVCISATAGKPEEFAKLSDEAPSNDQAMYESIQRYRRFVDGNRYNGGQQQQQQPKQWEVRPPDLSRDQRGNTKAQVEINKKGDNHDINAGWGKNINGPDSHKDTWHVGGSVRW (SEQ ID NO: 177) RlpA type I MKETTVVWAKLFLILIILAKPLGLKAVNECKRLGNNSCRSHGECCSGFCFIEPGWALGVCKRLGTPKKSDDSNNGKNIEKNNGVHERIDDVFERGVCSYYKGPSITANGDVFDENEMTAAHRTLPFNTMVKVEGMGTSVVVKINDRKTAADGKVMLLSRAAAESLNIDENTGPVQCQLKFVLDGSGCTPDYGDTCVLH HECCSQNCFREMFSDKGFCLPK (SEQ ID NO: 192)

In some cases, BRP modifies the growth and/or activity of one or more bacteria residing in the host bacteriocyte. In some cases, a BRP can be bioengineered to modulate its biological activity (eg, increase, decrease, or regulate) or to define its target microorganism. In some cases, BRP is produced by the translational machinery (e.g., ribosome, etc.) of the cell. In some cases, BRP is synthesized chemically. In some cases, BRP is formed from a polypeptide precursor. The precursor polypeptide may be cleaved (eg, processed by a protease) to form the BRP polypeptide itself. Thus, in some cases, BRP is produced from a precursor polypeptide. In some cases, the BRP includes a polypeptide that has undergone post-translational modifications, such as cleavage or the addition of one or more functional groups.

The functionally active BRP variants described herein are also useful in the compositions and methods described herein. In some cases, the BRP option is at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83 %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical , for example, in a specific region or in the entire sequence, the sequence of a BRP described herein, or a naturally occurring BRP.

The BRP described herein may be formulated for any of the routes of use described herein. The compositions disclosed herein may contain any number or type (e.g., classes) of BRP, such as at least about any number of 1 BRP, 2, 3, 4, 5, 10, 15, 20, or more BRP . The appropriate concentration of each BRP in the composition depends on factors such as the effectiveness, stability of the BRP, the number of different BRPs, the composition and methods of use of the composition. In some cases, the concentration of each BRP in the liquid composition is from about 0.1 ng/ml to about 100 mg/ml. In some cases, the concentration of each BRP in the solid composition is from about 0.1 ng/g to about 100 mg/g. In some cases, if the composition contains at least two types of BRP, the concentration of each type of BRP may be the same or different.

A modulatory agent comprising BRP described herein can be brought into contact with the target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) concentration of BRP within target host; (b) achieving a target level (eg, a predetermined or threshold level) of BRP concentration within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of BRP concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/and (e) modulating the fitness of the target host.

iii. Small molecules

Numerous small molecules (eg, antibiotic or metabolite) can be used in the compositions and methods described herein. In some cases, an effective concentration of any small molecule described herein can alter the level, activity or metabolism of one or more microorganisms (described above) resident in the host, the change resulting in increased fitness of the host.

A modulatory agent comprising a small molecule described herein may be brought into contact with a target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) of the low concentration molecules within the target host; (b) achieving a target level (eg, a predetermined or threshold level) of concentration of the small molecule within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of concentration of the small molecule within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/and (e) modulating the fitness of the target host.

The small molecules discussed later in this document, namely antibiotics and secondary metabolites, can be used to alter the levels, activity, or metabolism of target microorganisms, as discussed in the sections regarding enhancing the fitness of insects such as honeybees and silkworms.

(a) Antibiotics

The modulatory agent described herein may include an antibiotic. Any antibiotic known in the art can be used. Antibiotics are usually classified based on their mechanism of action, chemical structure, or spectrum of activity.

The antibiotic described herein can specifically target any function or growth process of bacteria and can be either bacteriostatic (eg, slow or prevent the growth of bacteria) or bactericidal (eg, kill bacteria). In some cases, the antibiotic is a bactericidal antibiotic. In some cases, a bactericidal antibiotic is an antibiotic that specifically targets the bacterial cell wall (eg, penicillins and cephalosporins); an antibiotic that specifically targets the cell membrane (for example, polymyxins); or an antibiotic that inhibits essential bacterial enzymes (eg, rifamycins, lipiarmycins, quinolones and sulfonamides). In some cases, the bactericidal antibiotic is an aminoglycoside. In some cases, the antibiotic is a bacteriostatic antibiotic. In some cases, a bacteriostatic antibiotic specifically targets protein synthesis (for example, macrolides, lincosamides and tetracyclines). Additional classes of antibiotics that may be used herein include cyclic lipopeptides (such as daptomycin), glycylcyclines (such as tigecycline), oxazolidinones (such as linezolid) or lipiarmycins (such as fidaxomicin). Examples of antibiotics include rifampicin, ciprofloxacin, doxycycline, ampicillin, and polymyxin B. Non-limiting examples of antibiotics are found in Table 9.

Table 9. Examples of antibiotics

Antibiotics Action Penicillins, cephalosporins, vancomycin Cell wall synthesis Polymyxin, gramicidin Membrane active agent, destroys the cell membrane Tetracyclines, macrolides, chloramphenicol, clindamycin, spectinomycin Inhibit protein synthesis Sulfonamides Inhibits folate-dependent pathways Ciprofloxacin Inhibits DNA gyrase Isoniazid, rifampicin, pyrazinamide, ethambutol, (miambutol), streptomycin Antimycobacterial agents

The antibiotic described herein may have any level of target specificity (eg, narrow or broad spectrum). In some cases, the antibiotic is a narrow-spectrum antibiotic and thus specifically targets specific types of bacteria, such as gram-negative or gram-positive bacteria. Alternatively, the antibiotic may be a broad-spectrum antibiotic that specifically targets a wide range of bacteria.

The antibiotics described herein may be formulated for any of the routes of administration described herein. The compositions disclosed herein may contain any number or type (e.g., classes) of antibiotics, such as at least about any number of 1 antibiotic, 2, 3, 4, 5, 10, 15, 20, or more antibiotics (eg, a combination of rifampicin and doxycycline or a combination of ampicillin and rifampicin). The appropriate concentration of each antibiotic in the composition depends on factors such as potency, stability of the antibiotic, amounts of different antibiotics, composition, and methods of administration of the composition. In some cases, if the composition contains at least two types of antibiotics, the concentration of each type of antibiotic may be the same or different.

The modulating agent, including an antibiotic described herein, can be brought into contact with the target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) concentration of the antibiotic within target host; (b) achieving a target level (eg, a predetermined or threshold level) of antibiotic concentration within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of antibiotic concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/or (e) modulating the fitness of the target host.

(b) Secondary metabolites

In some cases, the modulating agent in the compositions and methods described herein includes a secondary metabolite. Secondary metabolites are formed from organic molecules produced by the body. Secondary metabolites can act (i) as competitive agents used against bacteria, fungi, amoebae, plants, insects and large animals; (ii) as means of transporting metals; (iii) as agents of symbiosis between microorganisms and plants, nematodes, insects and larger animals; (iv) as sex hormones and (v) as differentiation effectors. Non-limiting examples of secondary metabolites are found in Table 10.

Table 10. Examples of secondary metabolites

Phenylpropanoids Alkaloids Terpenoids Quinones Steroids Polyketides Anthocyanins Acridines Carotenes Anthroquinones Heartfelt Erythromycin Coumarins Betalains Monoterpenes Benzoquinones glycosides Lovastatin and other statins Flavonoids Quinolizidines Sesquiterpenes Naphthoquinones Derivatives Discodermolide Derivatives Furonoquinones Diterpenes pregnenolone Aflatoxin B1 hydroxycinnamoyl Harringtonines Triterpenes Avermectins Isoflavonoids Isoquinolines Nystatin Lignans Indoles Rifamycin Phenolenones Purines Proanthocyanidins Pyridines Stilbenes Tropanes Tannins alkaloids

A secondary metabolite as used herein may include a metabolite from any known group of secondary metabolites. For example, secondary metabolites can be classified into the following groups: alkaloids, terpenoids, flavonoids, glycosides, natural phenols (e.g. gossypoacetic acid), enals (e.g. trans-cinnamaldehyde), phenazines, biphenols and dibenzofurans, polyketides, fatty acid synthase peptides, non-ribosomal peptides, peptides synthesized in ribosomes and subject to post-translational modifications, polyphenols, polysaccharides (eg chitosan) and biopolymers. For an in-depth review of secondary metabolites, see, for example, Vining, Annu. Rev. Microbiol. 44:395-427, 1990.

Secondary metabolites useful in the compositions and methods described herein include metabolites that alter the natural function of an endosymbiont (eg, a primary or secondary endosymbiont), bacteriocyte, or extracellular symbiont. In some cases, one or more of the secondary metabolites described herein are isolated as a result of high throughput screening (HTS) of antimicrobial compounds. For example, the HTS screen identified 49 antibacterial extracts that were specific for Gram-positive and Gram-negative bacteria from over 39,000 crude extracts from organisms growing in different ecosystems in one specific region. In some cases, the secondary metabolite is transported inside the bacteriocyte.

In some cases, the small molecule is an inhibitor of vitamin synthesis. In some cases, the vitamin synthesis inhibitor is an analogue of the vitamin precursor. In certain cases, the vitamin precursor analogue is pantothenol.

In some cases, the small molecule is an analogue of an amino acid. In certain cases, the amino acid analogue is L-canavanine, D-arginine, D-valine, D-methionine, D-phenylalanine, D-histidine, D-tryptophan, D-threonine, D-leucine, L-NG-nitroarginine, or their combination.

In some cases, the small molecule is a natural antimicrobial compound such as propionic acid, levulinic acid, trans-cinnamaldehyde, nisin, or low molecular weight chitosan. The secondary metabolite described herein may be formulated for any of the routes of administration described herein. The compositions disclosed herein may contain any number or type (e.g., classes) of secondary metabolites, such as at least about any number of 1 secondary metabolite, 2, 3, 4, 5, 10, 15, 20, or more secondary metabolites. The appropriate concentration of each secondary metabolite in the composition depends on factors such as potency, stability of the secondary metabolite, the amount of different secondary metabolites, composition and methods of use of the composition. In some cases, if the composition contains at least two types of secondary metabolites, the concentration of each type of secondary metabolite may be the same or different.

A modulatory agent comprising a secondary metabolite described herein may be brought into contact with the target host in an amount and for a time sufficient to: (a) achieve a target level (e.g., a predetermined or threshold level) concentration of the secondary metabolite within the target host; (b) achieving a target level (eg, a predetermined or threshold level) of the concentration of the secondary metabolite within the intestine of the target host; (c) achieving a target level (eg, a predetermined or threshold level) of secondary metabolite concentration within the target host bacteriocyte; (d) modulating the level or activity of one or more microorganisms (eg, endosymbionts) in the target host; and/and (e) modulating the fitness of the target host.

iv. Bacteria as modulating agents

In some cases, the modulating agent described herein includes one or more bacteria. Numerous bacteria are useful in the compositions and methods described herein. In some cases, the agent is a species of bacteria found endogenously in the host. In some cases, the bacterial modulator is a species of endosymbiotic bacteria. Non-limiting examples of bacteria that may be used as modulating agents include all of the bacterial species described herein in Section II of the Detailed Description and the bacterial species listed in Table 1 starting on page 15. For example, the modulating agent may be a bacterial species of any type of bacteria present in the guts of insects, including Gammaproteobacteria, Alphaproteobacteria, Betaproteobacteria, Bacteroidetes, Firmicutes (such as Lactobacillus and Bacillus spp.), Clostridia, Actinomycetes, Spirochetes, Verrucomicrobia , and Actinobacteria .

In some cases, the modulator is a bacterium that promotes microbial diversity or otherwise modifies the host microbiota in a beneficial manner. In one case, bacteria may be provided to promote the development of the microbiome in honey bees. For example, the modulating agent may include, for example, Bartonella apis, Parasaccharibacter apium, Frischella perrara, Snodgrassella alvi, Gilliamela apicola, Bifidobacterium spp or Lactobacillus spp .

The bacterial modulatory agents discussed herein can be used to alter the level, activity, or metabolism of target microorganisms, as discussed in the sections regarding enhancing the fitness of insects such as honeybees and silkworms.

In some cases, such bacterial modulators are bacteria capable of breaking down the pesticides set forth in Table 12, including insecticides. Such insecticides include neonicotinoids such as imidacloprid or organophosphate insecticides such as fenitrothion. In some cases, pesticide-metabolizing bacteria are present at a concentration of at least 100,000 cells/ml (e.g., at least about 100,000 cells/ml, at least about 150,000 cells/ml, at least about 200,000 cells/ml, at least at least about 250,000 cells/ml, at least about 300,000 cells/ml, at least about 350,000 cells/ml, at least about 400,000 cells/ml, at least about 450,000 cells/ml or at least about 500,000 cells/ ml).

Examples 1-3, 5 and 6 describe how imidacloprid and fenitrothion degrading microorganisms can be identified and can then be used as modulators in insect hosts such as honey bees, giving the treated insect hosts a competitive advantage. It will be understood that altering the level, activity or metabolism of one or more host microorganisms includes the introduction of such pesticide-degrading microorganisms, such as imidacloprid-degrading or fenitrothion-degrading microorganisms, to insect hosts such as honey bees.

In some cases, such bacterial modulators are bacteria capable of producing nutrients, including amino acids (eg, methionine or glutamate). The nutrient-producing bacteria may be naturally occurring bacteria, for example, naturally occurring bacteria exogenous to the insect host. Such bacteria may be isolated from a population of bacteria, such as a population found in an environmental sample. Bacteria can be isolated that produce one or more amino acids in a manner that increases the production of amino acids in the host compared to a host that has not been introduced to the amino acid-producing bacteria. Amino acids that can be produced by bacteria in the host include methionine, alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glutamate, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine , threonine, tryptophan, tyrosine or valine. In some cases, the amino acid-producing bacteria are methionine-producing bacteria.

In some cases, nutrient-producing bacteria (e.g., amino acid-producing bacteria, e.g., methionine-producing bacteria) are present at a concentration of at least 100,000 cells/mL (e.g., at least about 100,000 cells/mL, at least about 150,000 cells/ml, at least about 200,000 cells/ml, at least about 250,000 cells/ml, at least about 300,000 cells/ml, at least about 350,000 cells/ml, at least about 400,000 cells/ml, according to at least about 450,000 cells/ml or at least about 500,000 cells/ml).

Examples 8, 9 and 10 describe how methionine-producing microorganisms can be identified which can then be used as modulators in insect hosts such as honey bees or in the Drosophila model organism to enhance host fitness (e.g. increase in amino acid content (for example, methionine content or glutamate content)).

v. Modifications regarding modulating agents

(a) Merger products

Any of the modulatory agents described herein may be fused or associated with an additional moiety. In some cases, the modulating agent includes a fusion product of one or more additional fragments (eg, 1 additional fragment, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more additional fragments). In some cases, the additional fragment is any of the modulatory agents described herein (eg, a peptide, polypeptide, small molecule, or antibiotic). Alternatively, the additional fragment may not act as a modulator per se, but may instead serve a secondary function. For example, the additional moiety may facilitate access to, binding to, or activation of the modulatory agent at a target site in the host (eg, host gut or host bacteriocyte) or in a target microorganism residing in the host (eg, honey bee or silkworm).

In some cases, the additional moiety may facilitate penetration of the modulating agent into the target host cell or target microorganism residing in the host. For example, the additional fragment may include a cell penetrating peptide. Cell penetrating peptides (CPPs) can be naturally occurring sequences derived from proteins; chimeric peptides, which are formed as a result of the fusion of two natural sequences; or synthetic CPPs, which are synthetically engineered sequences based on structure-activity relationship studies. In some cases, CPPs are characterized by the ability to widely penetrate cell membranes (eg, prokaryotic and eukaryotic cell membranes) with limited toxicity. In addition, CPPs may be characterized by the ability to penetrate cell membranes through energy-dependent and/or energy-independent mechanisms without the need for chiral recognition by specific receptors. CPPs may be coupled to any of the modulatory agents described herein. For example, CPP can bind to an antimicrobial peptide (AMP), such as a scorpion peptide, such as UY192, fused to a cell penetrating peptide (eg, YGRKKRRQRRRFLSTIWNGIKGLLFAM; SEQ ID NO: 198). Non-limiting examples of CPPs are provided in Table 11.

Table 11. Examples of cell penetrating peptides (CPPs)

Peptide Origin Subsequence Derived from proteins rit Antennapedia RQIKIWFQNRRMKWKK (SEQ ID NO: 178) Peptide Tat Tat GRKKRRQRRRPPQ (SEQ ID NO: 179) pVEC Cadherin LLIILRRRIRKQAHAHSK (SEQ ID NO: 180) Chimera Transportan Galanin/Mastoparan GWTLNSAGYLLGKINLKALAALAKKIL (SEQ ID NO: 181) MPG HIV-gp41/T-antigen SV40 GALFLGFLGAAGSTMGAWSQPKKKRKV (SEQ ID NO: 182) Pep-1 HIV/T antigen SV40 reverse transcriptase KETWWETWWTEWSQPKKKRKV (SEQ ID NO: 183) Synthetic Polyarginines Based on Tat peptide (R) _{n ;} 6 < n < 12 MAP de novo KLALKLALKALKAALKLA (SEQ ID NO: 184) R ₆ W ₃ Based on penetratin RRWWRRWRR (SEQ ID NO: 185)

In other cases, the additional moiety facilitates binding of the modulating agent to the target microorganism (eg, fungus or bacterium) residing in the host. The additional fragment may include one or more targeting domains. In some cases, the targeting domain may target the modulating agent to one or more microorganisms (eg, bacteria or fungi) residing in the host's gut. In some cases, the targeting domain may target the modulating agent to a specific area in the host body (eg, the gut or host bacteriocyte) to access microorganisms that are normally present in the specified area of the host body. For example, the targeting domain may target the modulating agent to the foregut, midgut, or hindgut of the host. In other cases, the targeting domain may target the modulating agent to a bacteriocyte in the host and/or one or more specific bacteria residing in the host bacteriocyte. For example, the targeting domain may be a Galanthus nivalis lectin or agglutinin (GNA) linked to a modulatory agent described herein, for example an AMP, for example a scorpion peptide, for example Uy192.

(b) Pre- or prodomains

In some cases, the modulating agent may include a pre- or pro-amino acid sequence. For example, the modulating agent may be an inactive protein or peptide that can be activated by cleavage or post-translational modification of a pre- or prosequence. In some cases, the modulating agent is designed with an inactivating pre- or prosequence. For example, the pre- or prosequence may cover an activation site on a modulating agent, such as a receptor binding site, or may induce a conformational change in the modulating agent. Thus, upon cleavage of the pre- or prosequence, the modulating agent is activated.

Alternatively, the modulating agent may include a small pre- or pro-molecule, such as an antibiotic. The modulating agent may be an inactive small molecule described herein that can be activated in a target environment within the host. For example, a small molecule can be activated when a certain pH value is reached in the host's gut.

(c) Linkers

In cases where the modulating agent is linked to an additional moiety, the modulating agent may further comprise a linker. For example, the linker may be a chemical bond, such as one or more covalent bonds or non-covalent bonds. In some cases, the linker may be a peptide linker (eg, 2, 3, 4, 5, 6, 8, 10, 12, 14, 16, 20, 25, 30, 35, 40 or more amino acids in length). The linker may include any of the flexible, rigid, or cleavable linkers described herein.

The flexible peptide linker may include any of the linkers commonly used in the art, including linkers having sequences containing primarily Gly and Ser residues ("GS" linker). Flexible linkers may be suitable for connecting domains that require a certain degree of mobility or interaction, and may contain small, non-polar (eg, Gly) or polar (eg, Ser or Thr) amino acids.

Alternatively, the peptide linker may be a rigid linker. Rigid linkers are useful for maintaining a fixed distance between fragments and maintaining their independent functions. Rigid linkers may also be suitable if spatial separation of the domains is important to maintain the stability or biological activity of one or more components in the fusion product. Rigid linkers, for example, may have an alpha helical structure or a Pro-rich sequence, (XP) _n , with X being any amino acid, preferably Ala, Lys or Glu.

In a few other cases, the peptide linker may be a cleavable linker. In some cases, linkers may be cleaved under specific conditions, such as the presence of reducing reagents or proteases. In vivo cleavable linkers can exploit the reversible nature of the disulfide bond. One example involves a thrombin-sensitive sequence (eg, PRS) between two Cys residues . Thrombin treatment of CPRSC in vitro results in cleavage of the thrombin-sensitive sequence while the reversible disulfide bond remains intact. Such linkers are known and described, for example, in Chen et al., Adv. Drug Deliv. Rev. 65(10):1357–1369, 2013. Cleavage of linkers in fusion products can also be carried out by proteases that are expressed in vivo in specific cells or tissues of the host or microorganisms living in the host. In some cases, upon reaching the target site or cell, cleavage of the linker may release a free functional modulating agent.

The fusion products described herein may alternatively be associated with a linking molecule, including a hydrophobic linker such as a negatively charged sulfonate group; lipids, such as poly(--CH2--) hydrocarbon chains, such as a polyethylene glycol (PEG) group, unsaturated variants thereof, hydroxylated variants thereof, amidated or otherwise N-containing variants thereof, non-carbon linkers; carbohydrate linkers; phosphodiester linkers or other molecules capable of covalently linking two or more molecules, for example, two modulating agents. Non-covalent linkers can be used, such as hydrophobic lipid globules to which the modulating agent binds, for example through a hydrophobic region of the modulating agent or a hydrophobic extension of the modulating agent, such as a series of residues rich in leucine, isoleucine, valine or possibly also alanine, phenylalanine or even tyrosine, methionine, glycine or other hydrophobic residue. The modulatory agent may be coupled by chemical charge interaction such that a positively charged moiety of the modulatory agent is bound to a negatively charged other modulatory agent or additional moiety.

IV. Formulations and compositions

The compositions described herein may be formulated in pure form (eg, the composition contains only a modulating agent) or together with one or more additional agents (such as an excipient, delivery vehicle, carrier, diluent, stabilizer, etc.) to facilitating the use or delivery of the compositions. Examples of suitable excipients and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline, syrup , methylcellulose, methyl and propyl hydroxybenzoates, talc, magnesium stearate and mineral oil.

In some cases, the composition contains a delivery vehicle or carrier. In some cases, the delivery vehicle includes a filler. Exemplary excipients include, but are not limited to, solid or liquid carrier materials, solvents, stabilizers, sustained release formulation excipients, colorants, and surfactants. In some cases, the delivery vehicle is a stabilizing delivery vehicle. In some cases, the stabilizing delivery vehicle includes a stabilizing excipient. Exemplary stabilizing excipients include, but are not limited to, epoxidized vegetable oils, antifoam agents such as silicone oil, preservatives, viscosity regulators, binders, and tackifiers. In some cases, the stabilizing delivery vehicle is a buffer suitable for the modulating agent. In some cases, the composition is microencapsulated in a polymer bead-based delivery vehicle. In some cases, the stabilizing delivery vehicle protects the modulating agent from UV and/or acidic conditions. In some cases, the delivery vehicle contains a pH buffer. In some cases, the composition is formulated such that it has a pH in the range from about 4.5 to about 9.0, including, for example, any of the pH ranges from about 5.0 to about 8.0, from about 6.5 to about 7.5 or about 6.5 to about 7.0.

Depending on the intended purposes and prevailing circumstances, the composition may be formulated as emulsion concentrates, suspension concentrates, direct spray or dilute solutions, coating pastes, diluted emulsions, sprayable powders, soluble powders, dispersible powders, wettable powders, dust preparations , granules, forms encapsulated in polymeric substances, microcapsules, foams, aerosols, carbon dioxide gas preparations, tablets, resin-based preparations, paper-based preparations, non-woven fabric-based preparations or knitted or woven fabric-based preparations. In some cases, the composition is a liquid. In some cases, the composition is a solid. In some cases, the composition is an aerosol, such as in a pressurized aerosol can. In some cases, the composition is present in the waste (such as excrement) of the pest. In some cases, the composition is present inside or on the body surface of the living pest.

In some cases, the delivery vehicle is food or water for the host. In other cases, the delivery vehicle provides a food source for the host. In some cases, the delivery vehicle is a food bait for the host. In some cases, the composition is an edible product that is ingested by the host. In some cases, the composition is delivered by a host to a second host and is absorbed by the second host. In some cases, the composition is absorbed by the host or co-host, and the composition is released into the environment of the host or co-host with waste (such as excrement) of the host or co-host. In some cases, the modulating agent is included in a food bait intended for absorption by the host or transport back to its colony.

In some cases, the modulating agent may comprise from about 0.1% to about 100% of the composition, such as any amount from about 0.01% to about 100%, from about 1% to about 99.9%, from about 0.1% to about 10%, from about 1% to about 25%, from about 10% to about 50%, from about 50% to about 99%, or from about 0.1% to about 90% of the active ingredients (such such as phage, lysine or bacteriocin). In some cases, the composition contains at least any amount of 0.1%, 0.5%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% , 90%, 95% or more active ingredients (such as phage, lysine and bacteriocin). In some cases, concentrated products are preferred as commercial products, and the end user typically uses diluted products that have a significantly lower concentration of the active ingredient.

Any of the formulations described herein can be applied in the form of a bait, coil, electric mat, fuming preparation, fumigant or sheet.

i. Liquid formulations

The compositions provided herein may be in the form of a liquid composition. Liquid formulations are typically mixed with water, but in some cases they can be used with vegetable oil, diesel fuel, kerosene or other light oil as a carrier. The amount of active ingredient often ranges from about 0.5 to about 80 percent by weight.

The emulsion concentrate formulation may contain a liquid active ingredient, one or more petroleum-based solvents, and an agent that causes the formulation to mix with water to form an emulsion. Such concentrates can be used in formulations for agriculture, ornamentals, turfgrass, forestry, building treatment, food processing, livestock and public health pests. They can be adapted for application equipment from small portable sprayers to hydraulic sprayers, low volume ground sprayers, fan foggers and low volume aerial sprayers. Some active ingredients dissolve easily in the liquid carrier. When mixed with a carrier, they form a solution in which no precipitation or separation occurs, such as a homogeneous solution. Formulations of this type may contain an active ingredient, a carrier and one or more other ingredients. Solutions can be used in any type of sprayer, indoors or outdoors.

In some cases, the composition may be formulated as an inverse emulsion. An inverse emulsion is a water-soluble active ingredient dispersed in an oil carrier. Inverse emulsions require an emulsifier that helps mix the active ingredient with a large volume of a petroleum-based carrier, usually fuel oil. Inverse emulsions help reduce drift. With other formulations, some spray drift occurs as water droplets begin to evaporate before reaching the target surfaces; As a result, the droplets can become very small and lightweight. Because oil evaporates more slowly than water, the inverse emulsion droplets shrink less and more of the active ingredient reaches its target. The oil additionally helps reduce surface runoff and improves resistance to rain. It additionally acts as an adhesive, improving surface coverage and suction. Since the droplets are relatively large and heavy, it is difficult for them to penetrate the coating of the undersides of the leaves. Inverse emulsions are most widely used in rights-of-way where drift into susceptible non-target areas may be a problem.

The flowable or liquid formulation shares many of the characteristics of emulsion concentrates and wettable powders. Manufacturers use these formulations when the active ingredient is a solid that is neither water nor oil soluble. The active ingredient, impregnated with a substance such as clay, is ground to a very fine powder. The powder is then suspended in a small amount of liquid. The resulting liquid product is quite dense. Flowable and liquid formulations have many of the characteristics of emulsion concentrates, and they also have similar disadvantages. They require moderate shaking to maintain suspension and leave visible residues similar to those of wettable powders.

Flowable/liquid formulations are easy to handle and apply. Since they are liquids, they are subject to splashing and splashing. They contain solid particles, so they contribute to abrasive wear of nozzles and pumps. Flowable and liquid suspensions settle in their containers. Because pourable and liquid formulations are prone to settling, packaging in five-gallon or smaller containers makes remixing easier.

Aerosol formulations contain one or more active ingredients and a solvent. Most aerosols have a low percentage of active ingredients. There are two types of aerosol formulations - the ready-to-use type, widely available in pressurized, sealed containers, and products used in electric or gasoline-powered aerosol generators that release the formulation as smoke or mist.

Ready-to-use aerosol formulations are typically small, self-contained units that release the formulation when the nozzle valve is actuated. The composition is passed through a thin hole using an inert gas under pressure to form fine droplets. These products are used in greenhouses, small areas inside buildings, or confined areas outdoors. Commercial models that hold up to 5 pounds of active ingredient are usually refillable.

Smoke- or fog-forming aerosol compositions are not under pressure. They are used in machines that break a liquid formulation into a fine mist or dense mist (aerosol) using a rapidly rotating disk or heated surface.

ii. Dry or solid formulations

Dry formulations can be divided into two types: ready-to-use and concentrates, which must be mixed with water for use as a spray. Most dusted formulations are ready-to-use and have a low percentage of active ingredients (less than about 10 percent by weight) along with a very fine dry inert carrier made from talc, chalk, clay, nutshells or volcanic ash. The size of individual dust particles varies. Some dust formulations are concentrates and have a high percentage of active ingredients. Before use, they are mixed with dry inert carriers. Powders are always used dry and can easily drift to non-target areas.

iii. Granular or pelletized formulations

In some cases, the composition is formulated in the form of granules. Granular formulations are similar to dust formulations, except that the granular particles are larger and heavier. Coarse particles can be obtained from materials such as clay, corn cobs or walnut shells. The active ingredient coats the outer surface of the granules or is absorbed into them. The amount of active ingredient may be relatively small, typically ranging from about 0.5 to about 15 percent by weight. Granular formulations are most often used for application to soil, insects or soil-dwelling nematodes, or absorption by plants through roots. Granular formulations are sometimes applied by airplane or helicopter to minimize drift or penetration into dense brush. After use, the granules may slowly release the active ingredient. Some granules require soil moisture to release the active ingredient. Granular formulations are also used to control mosquito larvae and other aquatic pests. Granules are used in pest control operations in agriculture, building treatments, ornamentals, turfgrass, aquatic environments, rights-of-way, and public health (for stinging insects).

In some cases, the composition is formulated in the form of pellets. Most pelletized formulations are very similar to granular formulations; these terms are used interchangeably. However, in a pelletized formulation, all particles have the same mass and shape. Particle uniformity facilitates application using precision operating equipment.

iv. Powders

In some cases, the composition is formulated as a powder. In some cases, the composition is formulated as a wettable powder. Wettable powders are dry, finely divided formulations that appear as dust-like preparations. They usually need to be mixed with water to be used as a spray. However, few products can be used as a dust preparation or as a wettable powder - the choice is up to the consumer. Wettable powders have from about 1 to about 95 percent by weight active ingredient; in some cases more than about 50 percent. The particles do not dissolve in water. They settle quickly unless constantly agitated to keep them suspended. They can be used on most pest problems and on most types of spray equipment where agitation is possible. Wettable powders have excellent residual activity. Due to their physical properties, most of the composition remains on the surface of treated porous materials such as concrete, plaster and untreated wood. In such cases, only water penetrates the material.

In some cases, the composition is formulated as a soluble powder. Soluble powder formulations look like wettable powders. At the same time, when mixed with water, soluble powders dissolve easily and form a true solution. Once they are thoroughly mixed, no additional shaking is required. The amount of active ingredient in soluble powders ranges from about 15 to about 95 percent by weight; in some cases more than about 50 percent. Soluble powders have all the advantages of wettable powders and none of the disadvantages, except for the danger of inhalation during mixing.

In some cases, the composition is formulated as granules dispersible in water. Water-dispersible granules, also known as dry flowable formulations, are similar to wettable powders except that they are dust-like and are formulated into small, easily identifiable granules. Water dispersible granules must be mixed with water before use. Once in water, the granules break down into fine particles similar to wettable powders. The composition requires continuous agitation to maintain it suspended in water. The percentage of active ingredient is high, often at least 90 percent by weight. Water-dispersible granules have many of the same advantages and disadvantages of wettable powders, except that they are more easily defined and mixed. Due to their low dust particle content, they pose a lower inhalation hazard to the consumer during use.

v. Bait

In some cases, the composition includes bait. The bait may be in any suitable form, such as solid, paste, pellet or powder form. The bait may also be transferred by the host back into the population of said host (eg, colony or hive). The bait can then act as a food source for other members of the colony, thereby providing an effective modulator to a large number of hosts and potentially the entire host colony.

Baits may be provided in a suitable "house" or "trap". Such houses and traps are commercially available, and existing traps can be adapted to include the compositions described herein. The house or trap may be provided, for example, in the form of a box, and may be provided ready-made or may be made, for example, from folding cardboard. Suitable materials for the house or trap include plastic and cardboard, particularly corrugated cardboard. The internal surfaces of traps can be lined with a sticky substance to limit the movement of the host when entering the trap. The house or trap may contain a suitable groove inside which can hold the bait in place. A trap is different from a house because the owner cannot easily leave the trap once caught, whereas the house acts as a "feeder" that provides the owner with a preferred environment in which they can feed and feel safe from predators.

vi. Attractants

In some cases, the composition contains an attractant (eg, a chemoattractant). The attractant may attract an adult host or an immature host (eg, a larva) to a location nearby the composition. Attractants include pheromones, chemicals secreted by animals, particularly insects, that influence the behavior or development of other individuals of the same species. Other attractants include sugar and protein hydrolysate syrups, yeast and rotting meat. Attractants can also be combined with the active ingredient and sprayed onto leaves or other objects in the treated area.

Various attractants are known to influence host behavior, such as the host's search for food, oviposition or mating sites, or mating partners. Attractants useful in the methods and compositions described herein include, for example, eugenol, phenethylpropionate, ethyldimethylisobutylcyclopropanecarboxylate, propylbenzodioxanecarboxylate, cis-7,8-epoxy-2-methyloctadecane, trans-8,trans-0-dodecadienol, cis -9-tetradecenal (and cis-11-hexadecenal), trans-11-tetradecenal, cis-11-hexadecenal, (Z)-11,12-hexadecadienal, cis-7-dodecenyl acetate, cis-8-dodecenyl acetate, cis-9 -dodecenyl acetate, cis-9-tetradecenyl acetate, cis-11-tetradecenyl acetate, trans-11-tetradecenyl acetate (and cis-11), cis-9,trans-11-tetradecadienyl acetate (and cis-9,trans-12), cis-9 ,trans-12-tetradecadienyl acetate, cis-7,cis-11-hexadecadienyl acetate (and cis-7,trans-11), cis-3,cis-13-octadecadienyl acetate, trans-3,cis-13-octadecadienyl acetate, anethole and isoamyl salicylate .

Agents other than chemoattractants can also be used to attract insects, including lighting devices with different wavelengths or colors of radiation.

vii. Nanocapsules/microencapsulation/liposomes

In some cases, the composition is provided as a microencapsulated formulation. Microencapsulated formulations are mixed with water and sprayed in the same manner as other sprayable formulations. Once sprayed, the plastic coatings break down and slowly release the active ingredient.

viii. Carriers

Any of the compositions described herein may be formulated to include a modulating agent described herein and an inert carrier. Such a carrier may be a solid carrier, a liquid carrier, a gel carrier, and/or a gaseous carrier. In certain embodiments, the carrier may be a seed coating agent. A seed coating agent is any non-naturally occurring composition that adheres, in whole or in part, to the surface of the seed. The composition may further contain an excipient or a surfactant. The composition may also contain one or more modulating agents to broaden the spectrum of action.

The solid carrier used for the formulation includes fine clay powder or granules (such as kaolin clay, diatomaceous earth, bentonite clay, Fubasami clay, acid clay, etc.), synthetic hydrated silica, talc, ceramic materials, others inorganic materials (e.g. sericite, quartz, sulfur, activated carbon, calcium carbonate, hydrated silica, etc.), a substance that can be sublimated and exists in solid form at room temperature (e.g. 2,4,6- triisopropyl-1,3,5-trioxane, naphthalene, p-dichlorobenzene, camphor, adamantane, etc.); wool; silk; cotton; hemp; pulp; synthetic resins (for example, polyethylene resins such as low-density polyethylene, unmodified low-density polyethylene and high-density polyethylene; ethylene-vinyl ester copolymers, such as ethylene-vinyl acetate copolymers; ethylene-methacrylic acid ester copolymers, such as ethylene-vinyl ester copolymers methyl methacrylate and ethylene-ethyl methacrylate copolymers; ethylene-acrylic acid copolymers, such as ethylene-methyl acrylate copolymers and ethylene-ethyl acrylate copolymers; ethylene-vinylcarboxylic acid copolymers, such as ethylene-acrylic acid copolymers; ethylene-tetracyclododecene copolymers; polypropylene resins, such such as propylene homopolymers and propylene-ethylene copolymers; poly-4-methylpentene-1, polybutene-1, polybutadiene, polystyrene; acrylonitrile-styrene copolymer resins; styrenic elastomers, such as acrylonitrile-butadiene-styrene copolymer resins, block copolymers of conjugated dienes and styrene and hydrides of block copolymers of conjugated dienes and styrene; fluorine rubbers; acrylic resins such as poly(methyl methacrylate); polyamide resins such as nylon 6 and nylon 66; polyester resins such as polyethylene terephthalate, polyethylene naphthalate, polybutylene terephthalate and polycyclohexylene dimethylene terephthalate, polycarbonates, polyacetals, polyacrylsulfones, polyarylates, polyhydroxybenzoic acid polyesters, polyether imide, polyether carbonates, polyphenylene ether resins, polyvinyl chloride, polyvinylidene chloride , polyurethane and porous resins such as polyurethane foam, polypropylene foam or polyethylene foam, etc. etc.), glasses, metals, ceramic materials, fibrous materials, fabrics, knitted fabrics, sheets, paper, yarn, foam, porous substances and multifilaments.

The liquid carrier may include, for example, aromatic or aliphatic hydrocarbons (for example, xylene, toluene, alkylnaphthalene, phenylxyl ethane, kerosene, gas oil, hexane, cyclohexane, etc.), halogenated hydrocarbons (for example, chlorobenzene, dichloromethane, dichloroethane, trichloroethane etc.), alcohols (e.g. methanol, ethanol, isopropyl alcohol, butanol, hexanol, benzyl alcohol, ethylene glycol, etc.), ethers (e.g. diethyl ether, ethylene glycol dimethyl ether, diethylene glycol monomethyl ether, monoethyl ether diethylene glycol, propylene glycol monomethyl ether, tetrahydrofuran, dioxane, etc.), esters (e.g. ethyl acetate, butyl acetate, etc.), ketones (e.g. acetone, methyl ethyl ketone, methyl isobutyl ketone, cyclohexanone, etc.), nitriles (e.g. acetonitrile, isobutyronitrile, etc.), sulfoxides (e.g. dimethyl sulfoxide, etc.), amides (e.g. N,N-dimethylformamide, N,N-dimethylacetamide, cyclic imides (e.g. N-methylpyrrolidone) , alkylidene carbonates (such as propylene carbonate, etc.), vegetable oil (such as soybean oil, cottonseed oil, etc.), vegetable essential oils (such as orange oil, hyssop oil, lemon oil, etc.) or water.

The gaseous carrier may include, for example, butane gas, polytetrafluoroethylene gas, liquefied petroleum gas (LPG), dimethyl ether and carbon dioxide gas.

ix. Excipients

In some cases, the composition provided herein may contain an excipient. Excipients are chemicals that are inactive. Excipients are premixed into the formulation or added to the spray tank to enhance mixing or application or to enhance functional characteristics. They are widely used in products intended for use on leaves. Excipients can be used to customize the formulation to suit specific needs and adjust for local conditions. Excipients may be designed to perform specific functions, including wetting, spreading, adhesion, reducing evaporation, reducing volatilization, buffering, emulsifying, dispersing, reducing spray drift and reducing foaming. No single excipient can perform all of these functions on its own, but compatible excipients can often be combined to perform multiple functions simultaneously.

Non-limiting examples of excipients included in the composition include binders, dispersants and stabilizers, in particular, for example, casein, gelatin, polysaccharides (for example, starch, acacia, cellulose derivatives, alginic acid, etc.), derivatives lignin, bentonite, sugars, synthetic water-soluble polymers (e.g. polyvinyl alcohol, polyvinylpyrrolidone, polyacrylic acid, etc.), PAP (isopropyl acid phosphate), BHT (2,6-di-tert-butyl-4-methylphenol), BHA (a mixture of 2-tert-butyl-4-methoxyphenol and 3-tert-butyl-4-methoxyphenol), vegetable oils, mineral oils, fatty acids and fatty acid esters.

x. Surfactants

In some cases, the composition provided herein contains a surfactant. Surfactants, also called wetting agents and spreaders, physically change the surface tension of the spray droplet. In order for the composition to properly perform its function, a drop of spray must be able to wet the leaves and be distributed evenly over the leaf. Surfactants increase the area covered by the composition, thereby increasing the effect of the chemical on the pest. Surfactants are especially important when applying the formulation to waxy or hairy leaves. Without proper wetting and distribution, spray droplets often run off or fail to adequately cover leaf surfaces. At the same time, too much surfactant can cause excess runoff and reduce efficiency.

Surfactants are classified based on the way they ionize, or break down, into electrically charged atoms or molecules called ions. A surfactant with a negative charge is anionic. A surfactant with a positive charge is cationic, and a surfactant without an electrical charge is nonionic. The activity of a formulation in the presence of a nonionic surfactant may differ significantly from the activity in the presence of a cationic or anionic surfactant. Selecting the wrong surfactant can reduce the effectiveness of the pesticide product and harm the target plant. Anionic surfactants are most effective when used with contact pesticides (pesticides that control the pest through direct contact rather than systemic absorption). Cationic surfactants should never be used as stand-alone surfactants as they are usually phytotoxic.

Nonionic surfactants, often used with systemic pesticides, facilitate penetration of pesticide sprays through plant cuticles. Nonionic surfactants are compatible with most pesticides, and for most EPA-registered pesticides that require a surfactant, the nonionic type is recommended. Excipients include, but are not limited to, adhesives, dry extenders, plant penetration agents, compatibilizers, pH buffers or modifiers, additional drift control components, antifoams, and thickeners.

Non-limiting examples of surfactants included in the compositions described herein include alkyl sulfate ester salts, alkyl sulfonates, aclylaryl sulfonates, alkyl aryl ethers and polyoxyethylenated products thereof, polyethylene glycol ethers, polyhydric alcohol esters, and sugar alcohol derivatives.

xi. Combinations

In the compositions and in the applied forms obtained from these compositions, the modulating agent may be mixed with other active compounds, such as pesticides (for example, insecticides, sterilants, acaricides, nematicides, molluscicides or fungicides; see, for example, pesticides listed in Table 12), attractants, growth regulators or herbicides. As used herein, the term “pesticide” refers to any substance or mixture of substances intended to prevent, interfere with, repel or mitigate the effects of any pest. A pesticide may be a chemical or biological agent used against pests, including insects, shellfish, pathogens, weeds, nematodes, and microorganisms that compete with humans for food, destroy property, spread disease, or are a negative nuisance. The term "pesticide" may further cover other biologically active molecules such as antibiotics, antiviral pesticides, antifungals, anthelmintics, nutrients, pollen, sucrose and/or agents that stop or slow down insect movement.

In cases where the modulating agent is applied to plants, a mixture with other known compounds, such as herbicides, fertilizers, growth regulators, antidotes, chemical signal substances or other agents for improving the properties of the plant, is also possible.

V. Delivery

A host described herein may be exposed to any of the compositions described herein in any suitable manner that delivers or administers the composition to the insect. The modulating agent can be delivered alone or in combination with other active or inactive substances and can be applied, for example, by nebulization, microinjection, through plants, by irrigation, dipping, in the form of concentrated liquids, gels, solutions, suspensions, sprays, powders, pellets, briquettes, tiles, etc., formulated to deliver an effective concentration of the modulating agent.

The amounts and locations for application of the compositions described herein are generally determined by the host environment, the stage of the life cycle at which the host microorganisms can be targeted with a modulating agent, the site in which application is to be made, and the physical and functional characteristics of the modulating agent. The modulating agents described herein can be administered to the insect by oral ingestion, but can also be administered by means that penetrate the cuticle or enter the insect's respiratory system.

In some cases, the insect may simply be "wetted" or "sprayed" with a solution containing a modulating agent. Alternatively, the modulating agent may be associated with a food component (eg, edible) to the insect to facilitate delivery and/or to enhance uptake of the modulating agent by the insect. Methods of oral administration include, for example, directly mixing the modulatory agent with the insect's food, spraying the modulatory agent into the insect's habitat or field, and engineering-based approaches in which a species that is administered as food is engineered to express the modulatory agent. means, after which it is fed to the insect subject to negative influence. In some cases, for example, the modulating agent composition may be included in or topped with the insect's diet. For example, a composition based on a modulating agent can be sprayed onto a field of crops in which the insect lives.

In some cases, the composition is sprayed directly onto a plant, such as a crop, such as by backpack spraying, aerial spraying, spraying/pollinating crops, etc. In cases where the modulating agent is delivered to the plant, the plant receiving the modulating agent , can be at any stage of plant growth. For example, the formulated modulators can be used as a seed coating or root treatment during the early stages of plant growth, or as a total plant treatment during later stages of the crop cycle. In some cases, the modulating agent may be applied to the plant as a topical agent such that the host insect ingests or otherwise comes into contact with the plant upon interaction with the plant.

In addition, the modulating agent can be applied (for example, to the soil in which the plant grows, or to the water that is used to water the plant) as a systemic agent that is absorbed and distributed throughout the tissues (for example, stems or leaves) of the plant. host or host animal in such a way that the insect feeding on it will receive an effective dose of the modulating agent. In some cases, plants or organisms used as food may be genetically transformed to express a modulatory agent such that a host feeding on the plant or organism used as food ingests the modulatory agent.

Sustained or continuous release can also be achieved by coating the modulating agent or composition with modulating agent(s) with a soluble or biodegradable coating layer, such as gelatin, wherein the coating dissolves or degrades in the application environment, which then makes the modulating agent available, or by dispersing the agent in a soluble or degradable matrix. Such continuous release and/or distribution means that the devices can advantageously be used to sustainably maintain an effective concentration of one or more modulatory agents described herein in a particular host environment.

The modulating agent may also be included in the environment in which the insect grows, lives, reproduces, feeds, or infests. For example, the modulating agent may be included in a food container, feeder, protective wrap, or hive. In some applications, the modulating agent may be associated with a solid support for use in powder form or in a "trap" or "trough". As an example, in applications in which the composition is to be used as a trap or bait for a particular insect host, the compositions may also be bound to a solid support or encapsulated in a slow release material. For example, in cases where the host is a honey bee, the compositions described herein can be administered by delivering the composition to a honey bee hive or to at least one habitat where the honey bee grows, lives, reproduces or feeds.

VI. Screening

Included herein are screening assays to identify a modulating agent, wherein the modulating agent is effective in altering the host microbiota and thereby increasing host fitness (eg, insect or nematode fitness). For example, a screening assay can be used to identify one or more modulatory agents that specifically target specific microorganisms and/or specific hosts. In addition, screening assays can be used to identify one or more microorganisms with improved functional capabilities. For example, a screening assay may be effective in identifying one or more microorganisms with an improved ability to metabolize (e.g., break down) a pesticide (e.g., the pesticide listed in Table 12 or an insecticide known in the art, e.g., a neonicotinoid (e.g., imidacloprid) or an organophosphorus insecticide (e.g., a thiophosphate insecticide, e.g., fenitrothion)) or a plant allelochemical (e.g., caffeine, soybean cystatin N, monoterpenes, diterpene acids, or phenolic compounds). Delivery and colonization of the isolated microorganism in the host may increase the host's resistance to the pesticide (eg, the pesticide listed in Table 12) or plant allelochemical, thereby increasing host fitness. The methods may also be suitable for isolating microorganisms with improved ability to colonize any of the hosts described herein.

For example, to screen for microorganisms that increase host resistance to a pesticide (e.g., the pesticide listed in Table 12), a starter culture that contains the microorganisms (e.g., bacteria) and high concentrations of the pesticide (e.g., the pesticide listed in Table 12) can be used. or an insecticide known in the art, for example a neonicotinoid (eg imidacloprid) or an organophosphorus insecticide (eg a thiophosphate insecticide, eg fenitrothion)). In some cases, the pesticide may be provided in the form of an environmental sample spiked with the pesticide (eg, a soil sample). Alternatively, the pesticide (eg, the pesticide shown in Table 12) may be provided in pure form or in combination with other carriers. In addition, one or more isolates of microorganisms may be inoculated directly into the medium (eg, from a laboratory strain) or may represent an environmental sample containing one or more species of microorganisms. The growth medium can be liquid or solid. In some cases, the pesticide of interest is the only source of carbon or nitrogen for microorganisms in the environment. The culture may be subcultured (eg, inoculated into fresh medium with high levels of the pesticide) any number of times to enrich for and/or isolate microbial strains (eg, bacterial strains) capable of metabolizing the pesticide. The original crop or subcultures may be assessed using any methods known in the art to examine changes (eg, reduction) in pesticide levels in a sample (eg, HPLC). Isolates that reduce the concentration of a pesticide (e.g., the pesticide listed in Table 12, or a pesticide known in the art, e.g., a neonicotinoid (e.g., imidacloprid) or an organophosphorus insecticide (e.g., a thiophosphate insecticide, e.g., fenitrothion)) can be isolated for use as a modulating agent in any of the methods or compositions described herein.

The methods can be used to further select microorganisms described herein, including those isolated based on screening assays, with improved ability to colonize and survive in a host (eg, insect, eg, bee). For example, a bacterial isolate (eg, an isolate having the ability to degrade a pesticide) may be inoculated into the host. The host can then be examined at regular intervals for the presence of the bacterial isolate (eg, by culturing or isolating 16s RNA from the guts of the host (eg, honey bee)). Bacterial isolates that survive in the host (eg, the midgut of a honey bee) can be isolated for use as a modulating agent in any of the methods or compositions described herein.

Table 12. Pesticides Aclonifen Fenchlorazole-ethyl Pendimethalin Acetamiprid Phenothiocarb Penflufen Alanicarb Fenitrothion Penflufen Amidosulfuron Fenpropidine Pentachlorobenzene Aminocyclopyrachlor Fluazolate Penthiopyrad Amisulbrom Flufenoxuron Penthiopyrad Anthraquinone Flumetralin Pirimiphos-methyl Asulam, sodium salt Fluxapyroxad Prallethrin Benfuracarb Fuberidazole Profenofos Bensulide Glufosinate ammonium Proquinazid Beta-HCH; beta BCH Glyphosate Prothiophos Bioresmethrin Group: Borax, borate salts (see. Pyraclofos Blasticidin-S Group: Paraffin oils, mineral Pyrazacchlor Borax; disodium tetraborate Galfenprox Pyrazophos Boric acid Imiprotrine Pyridabene Bromoxynyl heptanoate Imidacloprid Pyridalil Bromoxynyl octanoate Ipconazole Pyridifention Carbosulfan Isopyrase Pyrifenox Chlorantraniliprole Isopyrase Quinmerac Chlordimeform Lenacil Rotenone Chlorfluazuron Magnesium phosphide Sedaksan Chlorprofam Metaflumizone Sedaksan Climbazole Metazachlor Silafluofen Clopyralid Metazachlor Syntofen Copper(II) hydroxide Metobromuron Spinetors Cyflufenamide Metoxuron Sulfoxaflor Cyhalothrin Metsulfuron-methyl Temefos Cyhalothrin, gamma Milbemectin Thiacloprid Decahydrate Naled Thiamethoxam Diafenthiuron Napropamide Tolfenpyrad Dimefuron Nicosulfuron Tralomethrine Dimoxystrobin Nitenpyram Tributyltin compounds Dinotefuran Nitrobenzene Tridifan Diquat dichloride o-phenylphenol Triflumizole Ditianon Oils Validamycin E-phosphamidon Oxadiargyl Zinc phosphide EPTC Oxycarboxin Etaboxam Paraffin oil Ethyrimol Penconazole

EXAMPLES

The following is an example of the methods of the present invention. It should be understood that various other embodiments may be practiced in light of the general description given above.

Example 1. Isolation of microorganisms that break down the nicotinoid imidacloprid

This example demonstrates the production of a library of microorganisms capable of degrading the neonicotinoid imidacloprid.

Experimental design

Four soil samples containing elevated imidacloprid concentrations were collected as described in Bonmatin et al. ( Environ. Sci. Pollut. Res. Int . 22:35–67, 2015) and diluted in Kauffman-Kearney minimal salt medium (MSM) to enrich for imidacloprid-degrading microorganisms. The three crop treatments for each soil sample included the following: 50 mL carbon-limited MSM with 83 mg/L imidacloprid as the sole carbon source; 50 ml nitrogen limited MSM with imidacloprid as the sole nitrogen source and sodium citrate and sucrose added as additional carbon sources; and 50 ml MSM broth with all ingredients added sodium citrate, sucrose and imidacloprid. Each culture was inoculated with 2 g of collected soil.

The cultures were kept on a shaker at 100 rpm and 27°C. Subcultures (1 ml of culture solution diluted in 25 ml of new medium) were prepared monthly to obtain enriched samples without the use of soil. Cultures were aliquoted periodically from shaker flasks for HPLC analysis as described in Tago et al. ( Microbes Environ . 21:58-64, 2006). For HPLC analysis, ≈7.0 × 10 ⁷ microorganisms were incubated in 1.0 ml of 20 mM sodium potassium phosphate buffer (pH 7.0) with 1.0 mM imidacloprid. After 20 minutes of incubation at 30°C, 1.0 ml of methanol was added. Imidacloprid and its partial degradation products were analyzed quantitatively using an HPLC system (600E pump and 2487 dual wavelength absorbance detector; Waters). The retention times and peak areas of the HPLC profiles were compared with those of known standards from Bonmatin et al. ( Environ. Sci. Pollut. Res. Int . 22:35-67, 2015).

Mixed enrichment cultures from soil that showed a decrease in imidacloprid in HPLC analysis upon incubation were surface plated using dilutions of 10 ^-1 to 10 ^-6 on nitrogen-limited agar plates with 36 mg/kg imidacloprid and streptomycin.

After incubation at 27°C for one week, the plates were screened for colonies that appeared morphologically different from each other (with differences in size). Individual colonies were plated onto new agar plates with imidacloprid as the sole nitrogen source. After three weeks, individual colonies were placed in 25 ml tryptic soy broth (TSB) with 25 mg/l imidacloprid.

After three days of growth, each of the isolates was centrifuged in Falcon conical tubes for 10 minutes at 6700 x g. The supernatant was removed and isolates were resuspended in sterile phosphate buffer to a total volume of 10 ml. A 2-ml sample of each isolate was inoculated into cultures in nitrogen-limited MSM and carbon-limited MSM; both cultures contained 30 mg/l imidacloprid in a total volume of 25 ml. Uninoculated controls were also prepared by inoculating 2 ml of phosphate buffer into each type of medium. All samples were wrapped in aluminum foil and placed on a shaker operating at 27°C and 100 rpm. The samples were then analyzed for imidacloprid concentration using HPLC.

After seven days of incubation, 25 ml of methanol was added to the flasks. Each sample was then sonicated for six minutes at 50% duty cycle and centrifuged at 6700 x g. The supernatant was then analyzed and imidacloprid was quantified by HPLC as described herein.

Isolates that reduced imidacloprid concentrations were stored at -80°C in a 50% glycerol solution.

Example 2: Selection of imidacloprid-degrading isolates that survive in the midgut of bees

This example demonstrates the possibility of selecting an imidacloprid-degrading microorganism from the isolates described in Example 1 that is capable of surviving in the bee midgut.

Treatment regimen The ten isolates described in Example 1 were treated at 0 (negative control), 10 ² , 10 ⁵ or 10 ⁸ CFU/ml in 10 ml of 50% sugar syrup.

Experimental design

Approximately fifty bees per bacterial isolate were collected and kept in a wooden cage. The bacterial strains isolated in the previous example were cultured and prepared for feeding as follows. Cells from each isolate were washed and diluted to OD600 0.1 with MSM supplemented with 50% (w/v) sugar syrup and incubated at 30°C for cell growth. Four concentrations of fresh bacterial cultures were administered to bees as food (0-10 ⁸ CFU/ml) as follows. Different concentrations of isolates were placed in a 15 ml tube with six small narrow holes into which the bees inserted their proboscis and ate solutions containing the isolate. Fresh cultures containing the isolate were prepared daily for use as feed, and bee mortality was assessed each day for the entire duration (7 days) of the experiment by counting and removing dead bees. After 7 days, the solution containing the isolate was replaced with 15% (w/v) sucrose for 48 hours.

Ten bees were removed from each replicate every 24 hours for bacterial analysis; the first sample was collected before the start of the experiment, and this was followed by sampling at seven additional time points. After each sample collection, bees were anesthetized on ice and the midgut was removed. Replicates for each time point were pooled and homogenized in 150 μl of 1× PBS, then serially diluted and plated on nitrogen-limited agar plates with 36 mg/kg imidacloprid to grow bacteria under selective growth conditions at 30°C for 2 days. .

Bacterial isolates that survived in the bee midgut were selected for administration to improve bee resistance to neonicotinoids.

Example 3. Increasing the resistance of bees to neonicotinoids by introducing bacteria that break down imidacloprid

This example demonstrates the possibility of increasing the resistance of bees to insecticides, and more specifically, to neonicotinoids. Bees play an important role in the pollination of flowering plants and are the main type of pollinator in many ecosystems containing flowering plants. It is estimated that one third of human food resources depend on pollination by insects, birds and bats, much of which is provided by bees, particularly the domestic European honey bee. The number of colonies maintained by beekeepers has decreased as a result of urbanization, systematic use of pesticides, and exposure to tracheal mites and microscopic mites of the genus Varroa. Due to direct consumption of contaminated nectar and pollen from insecticide-treated plants, neonicotinoids in insecticides negatively affect foraging, learning, and memory in worker bees. By introducing imidacloprid-degrading bacteria into their microbiome, bees are expected to increase their resistance to neonicotinoids in the form of improved behavior such as increased foraging, hygiene behavior and survival.

Treatment regimen The bacterial isolate selected in Example 2 and identified as surviving in the midgut of bees was formulated at 10 ⁸ organisms in 10 ml of 50% sugar syrup.

Experimental design

The isolate selected in example 2 was prepared at a concentration of 10 ⁸ cells/ml (see example 2) and placed in a 15 ml test tube with six small narrow holes into which the bees inserted their proboscis and ate the solution containing the isolate. Fresh cultures containing the isolate were prepared daily for use as feed. Pre-treatment lasted a total of seven days.

Approximately 1500, 3000, or 7000 worker bees from hives, estimated by the mass of Apis mellifera worker bees (0.5, 1, and 2 pounds, or 0.23, 0.45, and 0.1 kg, respectively), were pretreated with the sample isolate 2, prepared at a concentration of 10 ⁸ cells/ml and placed in a 15 ml test tube with six small narrow holes into which the bees inserted their proboscis and ate the solution containing the isolate. Fresh cultures containing the isolate were prepared daily for use as feed. Pre-treatment lasted a total of seven days.

After a 7-day pretreatment, each worker bee per hive was fed 2–4 grams of pollen supplement and sugar syrup (1:1) for 1–2 days before imidacloprid treatment. Smaller colonies (1500 and 3000 bees) were placed in observation hives with glass walls. Larger colonies (7000 bees) were placed in Ulster observation hives (Brushy Mountain Bee Supply, North Carolina), which contained a bottom box with 4 standard frames and a feeder with a divider board that provided syrup for treatment. All observation hives were placed under shelters and maintained at a constant temperature and relative humidity (23-25°C and 70%). In addition, all hives had an entrance leading to the outside world, which allowed the bees to independently forage for food in the surrounding agricultural fields and urban residential areas.

Once oviposition was confirmed visually, each colony was randomly assigned to imidacloprid treatment (0, 10, 20, 50, and 100 ppm) provided in 50% sugar syrup. The colonies were given proportional amounts of sucrose solution with imidacloprid through a dosing system similar to that described in example 2: 80, 160 and 320 ml for colonies of 1500, 3000 and 7000 bees, respectively. Syrup supplies were replenished every other day for 3 weeks. The amount of syrup was intended to supplement, but not support, the colonies, so the bees had to obtain their own food from other sources. Imidacloprid stock solutions (100 ppm) were prepared using 99.5 ± 0.5% technical grade imidacloprid purchased from Chem Service, Inc (PS-2086) dissolved by shaking in 50% sucrose overnight. Stock solutions were prepared every two weeks, and treatment solutions were prepared every week. Samples of treatment solutions (3-6 per dose) were selected at random and tested for residual concentrations to confirm dosage accuracy.

Measurements of behavioral phenotypes

Foraging activity was measured by recording the number of worker bees entering and exiting the colony through the entrance during one-minute observations twice daily. Hygiene behavior was used as a measure to assess the activity of worker bees in a hive and was defined as the ability of worker bees to identify and remove diseased and microscopic mite-infested brood, thereby limiting the transmission of pathogens and parasites within the colony. Hygienic behavior was determined using a freeze-killed brood assay in which a 3-inch (7.6 cm) polyvinyl chloride tube was carefully placed into a section of comb containing a large area of sealed pupal cells (taken from non-experimental field colonies). The number of empty cells was counted and recorded before pouring 400 ml of liquid nitrogen to freeze kill approximately 160 pupae. The frame was then temporarily placed in the bee colony, and after 24 hours the proportion of pupae completely or partially removed from the cells was quantified to assess hygienic behavior. Removal of freeze-killed brood was correlated with removal of diseased and parasitized brood.

Bees pre-treated with the bacterial isolate selected in Example 2 exhibited higher activity than bees treated with the control.

Example 4. Treatment of microscopic mites from the genus Varroa, which infect bees, with oxytetracycline solutions

This example demonstrates the possibility of killing or reducing the fitness of microscopic mites from the genus Varroa by treating them with an antibiotic solution. This example demonstrates that the effect of oxytetracycline on Varroa mites is mediated by altering populations of bacteria, such as Bacillus , endogenous to Varroa mites that are susceptible to oxytetracycline.

Microscopic mites of the genus Varroa are believed to be the primary cause of widespread colony collapse disorder (CCD), which is leading to massive declines in domestic honey bee Apis mellifera colonies worldwide. They attach themselves to the bees' abdomens and suck their blood, depriving them of nutrients and ultimately killing them. Although microscopic Varroa mites can be killed using chemically synthesized miteicides, these types of chemicals should be kept away from edible honey.

Therapy scheme. Oxytetracycline solution was formulated at 0 (negative control), 1, 10, or 50 μg/ml in 10 ml of sterile water with 0.5% sucrose and essential amino acids.

Experimental design

To determine whether adult Varroa mites during the breeding stage have a different susceptibility compared to phoretic microscopic mites or their offspring because their cuticle is not hardened, microscopic Varroa mites living on adult Apis mellifera bees and microscopic mites were collected. mites associated with larvae and pupae. This analysis examined the effects of antibiotic solutions on different types of microscopic mites and determined how their fitness is altered by targeting endogenous microorganisms such as Bacillus .

Microscopic brood mites were collected from honeycombs (or pieces of honeycomb) in colonies of bees infested with microscopic mites of the genus Varroa. Microscopic mites were collected from cells in which bee larvae develop.

Microscopic mites from the genus Varroa were divided into groups according to the age of their hosts in the brood and analyzed individually. The age of the brood was determined based on the morphological characteristics and pigmentation of the larvae or pupae as follows: microscopic mites of the genus Varroa, collected from curled larvae that were small enough to move around their cell, were placed in group 1; microscopic mites of the genus Varroa, collected from erect larvae that were too large to lie in the cell and began to extend straight with their mouthparts towards the opening of the cell, were placed in group 2; and microscopic mites of the genus Varroa collected from pupae were placed in group 3. Microscopic mites were kept on the larvae or pupae of their hosts in glass Petri dishes until 50 units were collected. This ensured that their standard nutritional conditions and physiological state remained unchanged. To prevent microscopic mites from separating from the larvae or pupae of their hosts or crawling on top of each other, only hosts at the same developmental stage were kept in the same dish.

The device - a stainless steel ring (inner diameter 56 mm, height 2-3 mm) and 2 glass circles (diameter 62 mm) - was cleaned with acetone and hexane or pentane to form a test arena. Oxytetracycline solutions and a control solution were applied to the device by uniformly spraying the glass disks and arena ring. To do this, 1 ml of solutions was loaded into the reservoir; set the distance from the surface to be sprayed to the bottom of the tube to 11 mm, and use a nozzle at 0.0275 inches. The pressure was adjusted (usually in the range of 350-500 hPa) until the amount of solution deposited was 1 ± 0.05 mg/cm ² . Antibiotic solutions were poured into appropriate dishes, covering the entire bottom of the dishes, and the residual liquid was evaporated under a fume hood. The ring was placed between glass circles to form a chamber. The chambers were used within 60 hours of preparation for a maximum of three assays to monitor the effects of antibiotic solutions on microscopic mites. 10 to 15 microscopic mites of the genus Varroa were introduced into this chamber, and the parts of the device were joined together by drops of melted wax. Microscopic mites collected from curled larvae, straightened larvae, white-eyed pupae and dark-eyed pupae with white and pale bodies were used.

After 4 hours, microscopic mites were transferred to a clean glass Petri dish (diameter 60 mm) with two or three white-eyed pupae (4-5 days after coating) for feeding. Microscopic mites were observed under a dissecting lens at 4 hours, 24 hours and 48 hours after treatment with antibiotic or control solutions and classified according to the following categories:

• Movable: They moved on their limbs, regardless of whether they were poked with a needle.

• Paralyzed: They moved one or more appendages, without stimulation or after stimulation, but they were unable to ambulate.

• Dead: motionless and unresponsive to 3 consecutive stimulations.

A sterile toothpick or needle was used to stimulate the microscopic mites by touching their limbs. New toothpicks or sterile needles were used to stimulate each group to avoid contamination between groups of microscopic mites.

Analyzes were carried out at 32.5°C and relative humidity 60-70%. If the mortality of control individuals exceeded 30%, then replication was excluded. Each experiment was repeated using four series of chambers.

Bacillus status in groups of microscopic mites from the genus Varroa was assessed using PCR. Total DNA was isolated from control (non-oxytetracycline-treated) and oxytetracycline-treated (whole body) individuals using a DNA isolation kit (OMEGA, Bio-tek) according to the manufacturer's protocol. Primers for Bacillus , forward primer 5'-GAGGTAGACGAAGCGACCTG-3' (SEQ ID NO: 186) and reverse primer 5'-TTCCCTCACGGTACTGGTTC-3' (SEQ ID NO: 187), were designed based on 23S-5S rRNA sequences obtained from the genome Bacillus (accession number: AP007209.1) (Takeno et al., J. Bacteriol. 194(17):4767–4768, 2012) using Primer 5.0 software (Primer-E Ltd., Plymouth, UK). PCR amplification cycles included an initial denaturation step at 95°C for 5 minutes, 35 cycles at 95°C for 1 minute, 59°C for 1 minute and 72°C for 2 minutes, and a final step elongation for 5 minutes at 72°C. Amplification products from oxytetracycline-treated and control samples were analyzed on 1% agarose gels, stained with SYBR Safe, and visualized using an imaging system.

Survival rates of microscopic Varroa mites treated with oxytetracycline solution were compared with microscopic Varroa mites treated with a negative control.

The survival rates and mobility of microscopic mites from the genus Varroa treated with oxytetracycline solution decreased compared to the control.

Example 5. Isolation of microorganisms that break down the organophosphate insecticide fenitrothion

This example demonstrates the production of a library of microorganisms capable of degrading the organophosphate insecticide fenitrothion.

Experimental design

Soil samples were obtained from various sites where fenitrothion had previously been used to control insects. Fenitrothion-degrading bacteria were isolated from soil samples as described (Microbes Environ. Vol. 21, No. 1, 58-64, 2006). Briefly, 1 g of soil sample was thoroughly mixed with 9 mL of sterile distilled water. Then the soil particles were centrifuged at RCF 1000 g for 5 minutes, and then 100 μl of the supernatant was inoculated into a medium with fenitrothion and mineral salts (0.4 g/l yeast extract, 0.4 g/l fenitrothion, 15 g/l bacteriological agar). The plates were cloudy when prepared because fenitrothion was an emulsion.

Colonies that formed clear zones around themselves and were likely to degrade or metabolize fenitrothion were isolated and regrown on LB agar, nutrient agar, glucose-yeast agar, TSA agar, BHI agar, or MRS agar. Once unique bacterial colonies were identified, their genomes were extracted using a genomic DNA extraction kit (Qiagen, DNeasy Blood and Tissue DNA Isolation Kit) according to the manufacturer's protocol.

The 16S rRNA gene was amplified using universal bacterial primers 27F (5'-AGAGTTTGATCMTGGCTCAG-3'; SEQ ID NO: 188) and 1492R (5'-TACCTTGTTACGACTT-3'; SEQ ID NO: 189) and using the following PCR conditions: 94°C for 2 minutes, 30 cycles of 94°C for 1 minute, 56°C for 1 minute and 72°C for 2 minutes, and a final extension at 72°C for 5 minutes. Gel electrophoresis was used to confirm that the amplicons were the correct size (~1500 bp), and products were excised from the gel and purified using a gel extraction kit (Qiagen, QIAquick) according to the manufacturer's protocol. Correctly sized amplicons were Sanger sequenced, and BLAST was used to align the sequence against various 16s rRNA gene sequence repositories for bacterial identification.

To determine whether the isolated bacterium degrades fenitrothion, ~10 ⁷ bacterial cells were incubated in 1 ml of 20 mM sodium potassium phosphate buffer (pH 7) with 1 mM fenitrothion, as described in PNAS, Vol. 109, No. 22, 8618-8622, 2012. After 30 minutes of incubation at 30°C, the reaction was stopped by adding an equal volume of methanol. Fenitrothion and its metabolite 3-methyl-4-nitrophenol were then analyzed by HPLC. The retention times and peak areas of the HPLC profiles were compared with those of known standards.

Unique bacterial isolates that had the ability to degrade fenitrothion were then stored frozen in glycerol at -80°C.

Example 6. Increasing the resistance of Drosophila melanogaster to fenitrothion by introducing bacteria that break down fenitrothion

This example demonstrates the ability to protect the Drosophila melanogaster insect model from one or more of the negative effects of insecticides in their diet, more specifically fenitrothion. The following approach is simulative for other insects, such as bees or other insects disclosed herein, for example, insects that are important pollinators of many cultivated flowering plants and vegetable plants. Many insecticides, including fenitrothion, have been shown to be toxic to bees.

Experimental design

Treatment regimen The bacterial isolates selected in Example 5 were composed of 10 ⁹ organisms in 100 μl of fly feeding medium with and without fenitrothion.

The medium used to feed the flies was a medium containing corn meal, molasses and yeast (11 g/l yeast, 54 g/l yellow corn meal, 5 g/l agar, 66 ml/l molasses and 4.8 ml l propionic acid). For experimental procedures, fenitrothion at 0, 10, and 100 ppm or phosphate-buffered saline as a negative control was infused into sterile fly feeding medium.

Development rate analysis

On the first day, 10 ⁹ fenitrothion-degrading bacteria described in Example 5 suspended in 100 μl of phosphate-buffered saline, or an equal volume of saline (negative control), were added to sterile fly feeding medium with or without fenitrothion. All samples were left to dry overnight at 25°C as described in Appl. Environ. Microbiol . Vol. 82, No. 20, 6204-6213, 2016.

On the second day, fertilized embryos collected from flies were treated with a 2% hypochlorite solution for 5 minutes and then washed with sterile water to remove any extracellular microorganisms from the embryos. 10 μl of a suspension of embryos in water (embryo:water suspension 1:3) was added to bacteria-inoculated or negative control fly food medium with or without fenitrothion. Cohorts with embryos on fly food medium were maintained at 25°C with a cycle of alternating 12 hours of light and 12 hours of darkness until the end of the experiment.

The time to puparium formation and the number of pupae produced were measured in each cohort. Time to adult emergence and emergence rate were measured in each sample. From the time the first adult emerged from the pupa, the number of emerging adult flies was counted every 12 hours and the rate of emergence was calculated.

Embryos on fly food medium infused with fenitrothion, inoculated with fenitrothion-degrading bacteria, were expected to develop faster than embryos on fly food medium infused with fenitrothion without fenitrothion-degrading bacteria.

Survival Analysis

Approximately 12 days before day 1, sterile embryos were obtained as previously described and raised on sterile fly food medium. Adults began to emerge from their pupae 11 days after embryo collection when reared on sterile fly food medium without fenitrothion at 25°C on a 12-hour light/12-hour dark cycle.

On the first day, 10 ⁹ fenitrothion-degrading bacteria in phosphate-buffered saline were added to sterile fly feeding medium as described in the previous example.

On day 2, 10 newly emerged sterile adult males and females were placed on bacteria-inoculated or negative control fly food medium with or without fenitrothion. The fly food medium was maintained at 25°C on a 12-h light/12-h dark cycle until the end of the experiment. The number of surviving male and female flies was counted every day until all the flies died. Survival assays were performed to evaluate the beneficial fitness effect produced by fenitrothion-degrading bacteria on fly survival.

Flies reared on fly food medium infused with fenitrothion and inoculated with fenitrothion-degrading bacteria were expected to survive longer than flies reared on fly food medium infused with fenitrothion without fenitrothion-degrading bacteria.

Example 7: Elimination of Entomopathogenic Bacteria from Drosophila melanogaster using Naturally Occurring Phages

This example demonstrates the possibility of eliminating bacterial insect pathogens (such as Serratia marcescens , Erwinia carotovora and Pseudomonas entomophila ) from Drosophila melanogaster using naturally occurring phages. This procedure may be suitable as a mock assay for bacterial elimination in other insect species such as bees.

Experimental design

Therapy scheme. We used collections of phage libraries containing the following phage families: Myoviridae, Siphoviridae, Podoviridae, Lipothrixviridae, Rudiviridae, Ampullaviridae, Bicaudaviridae, Clavaviridae, Corticoviridae, Cystoviridae, Fuselloviridae, Gluboloviridae, Guttaviridae, Inoviridae, Leviviridae, Microviridae, Plas maviridae, Tectiviridae.

Several environmental samples (soil, pond sediment and wastewater) were collected in sterile 1 L flasks over a period of 2 weeks and processed immediately after collection and then stored at 4°C. Solid samples were homogenized in sterile Luria broth (LB) or tryptic soy broth (TSB) double strength from Difco supplemented with 2 mM CaCl2 to a final volume of 100 ml. pH values and phosphate levels were measured using phosphate indicator strips. For purification, all samples were centrifuged at 3000-6000 g for 10-15 minutes at 4°C and filtered through 0.2 μm low protein binding filters to remove any remaining bacterial cells. The supernatant containing the phage library was then stored at 4°C in the presence of chloroform in a glass vial.

20-30 ml of phage library was diluted to a volume of 30-40 ml with LB broth. The target bacterial strain (e.g. , Serratia marcescens , Erwinia carotovora , and Pseudomonas entomophila ) was added (50–200 μl of culture grown overnight in LB broth) to enrich the population of phages that specifically target that particular bacterial strain in culture. This culture was incubated overnight at 37°C with shaking at 230 rpm. Bacteria from this enrichment culture were removed by centrifugation (3000-6000 g, 15-20 minutes, 4°C) and filtered (0.2 or 0.45 µm filter). 2.5 ml of bacteria-free culture was added to 2.5 ml of LB broth, and 50–100 μl of target bacteria was added back to the culture for further phage enrichment. The enrichment culture was grown overnight as described above. A sample from this enrichment culture was centrifuged at 13,000 g for 15 minutes at room temperature, and 10 μl of the supernatant was placed on a Petri dish of LB agar along with 100-300 μl of target bacteria and 3 ml of melted 0.7% soft agar. The dishes were incubated overnight at 37°C.

Each of the plaques observed in the bacterial lawn was collected and transferred to 500 μl of LB broth. A sample from this initial plaque culture was further plated onto the target bacteria. Plaque purification was performed three times for all detected phages to isolate one homogeneous phage from a heterogeneous mixture of phages.

Lysates from high titer phage plates (>1x 10^10 PFU/mL) were prepared by collecting the top layers from plates containing a host bacterial strain exhibiting confluent lysis. After adding 5 ml of buffer, the top soft agar layer was macerated, cleared by centrifugation and sterilized by filtration. The resulting lysates were stored at 4°C. High titer phage lysates were further purified by isopycnic centrifugation using CsCl as described in Summer et al., J . Bacteriol . 192:179-190, 2010.

DNA was isolated from CsCl-purified phage suspensions as described in Summer, Methods Mol. Biol. 502:27-46, 2009. A single isolated phage was sequenced as part of two pools of phage genomes using the 454 pyrosequencing method. Briefly, phage genomic DNA was mixed in equimolar amounts to a final concentration of approximately 100 ng/L. Pooled DNA was fragmented, ligated with a pool-specific multiplex identification tag (MID), and sequenced by pyrosequencing using a full-plate reaction on a sequencer (Roche) according to the manufacturer's protocols. Pooled phage DNA was present in two sequencing reactions. The output data corresponding to each of the pools was collected separately using software (454 Life Sciences), adjusting the settings so that only reads from one MID per assembly were included. The identity of individual contigs was determined by PCR using primers generated for the contig sequences and individual preparations of phage genomic DNA as a template. Sequencing software (Gene Codes Corporation) was used to assemble and edit sequences.

Chromosomal terminal structures of phages were determined experimentally. Cohesive ends (cos) for phages were determined by sequencing the ends of the phage genome and sequencing the PCR products obtained by ligation amplification of circularized genomic DNA as described in Summer, Methods Mol. Biol. 502:27–46, 2009. Protein coding regions were initially predicted using gene prediction software (Lukashin et al. Nucleic Acids Res. 26:1107–1115, 1998), purified by manual analysis in Artemis (Rutherford et al ., Bioinformatics 16:944-945, 2000) and analyzed by applying BLAST (E-value threshold 0.005) (Camacho et al., BMC Bioinformatics 10:421, 2009). Proteins of particular interest were further analyzed using sequence search software (Hunter et al., Nucleic Acids Res . 40:D306-D312, 2012).

Electron microscopy of CsCl purified phage (> 1×10^11 PFU/ml), which lysed pathogenicDrosophila types of bacteria, was performed by diluting the phage stock solution with a tryptic soy broth-based buffer. Phages were applied to fine carbon-coated 400 mesh grids, stained with 2% (wt/vol) uranyl acetate, and air dried. The samples were observed in a transmission electron microscope operating at an accelerating voltage of 100 kV. Five virions of each phage were measured to calculate means and standard deviations of capsid and appendage sizes as appropriate.

Incorporation of phages into food

The medium used to feed the flies was a medium containing corn meal, molasses and yeast (11 g/l yeast, 54 g/l yellow corn meal, 5 g/l agar, 66 ml/l molasses and 4.8 ml l propionic acid). Phage solutions were poured into the fly feeding medium to obtain final phage concentrations of ^0-108 PFU/ml.

S. marcescens , Erwinia carotovora and Pseudomonas entomophila bacteria were grown at 30°C in nutrient broth or LB broth.

Sterile fly embryos were obtained by treating fertilized embryos collected from flies with a 2% hypochlorite solution for 5 minutes and then rinsing with sterile water to remove any extracellular microorganisms. Fly larvae with target bacteria were obtained by inoculating 10 ⁹ CFU of bacteria onto sterile fly food medium and adding sterile fly embryos to this food medium. After 2 days, ten S. marcescens- infected first instar fly larvae with a range of phage concentrations ( ^0-108 PFU/ml) were added to the fly feeding medium. The larvae were left to grow on the food medium along with the phages for 3 days until they reached the third instar stage. Larvae were then collected and individually homogenized in nutrient broth or LB broth, plated on nutrient agar or LB agar plates, and incubated at 30°C. The number of CFU of S. marcescens obtained from larvae in fly food media containing different phage concentrations was recorded. This showed the number of live bacteria that were present in the flies' bodies.

It was expected that the number of live bacteria would be reduced in larvae raised on fly food media containing phages that counteract the bacteria.

Example 8. Creation of a library of natural microorganisms

This example demonstrates the isolation of bacteria from soil that naturally produce the amino acid methionine.

The medium used to isolate microorganisms is starch casein nitrate agar (10.0 g starch; 0.003 g casein; 0.02 g _KNO3 ; 0.02 g NaCl; 0.5 mg _MgSO4 ; 0.2 mg CaCO ₃ ; 0.1 mg FeSO ₄ ; 12.0 g agar; 1 l H ₂ O; pH 7.0) (Kuster and Williams, 1964). Each environmental soil sample (1.0 g) was suspended in 9 mL of sterile water, and 1 mL of the suspension was serially diluted tenfold with sterile distilled water. One milliliter of a 10 ^-5 dilution was inoculated into agar medium and incubated for 7 days at 30°C. At the end of this period, the plates were examined for growth. Individual white and dense colonies were collected and grown on new starch-casein-nitrate agar plates to create a library of isolates. After 7 days of growth at 30°C, the plates were kept at 4°C.

Example 9: Screening of Isolates that Produce Methionine

This example demonstrates a screening analysis of isolates from Example 8 that naturally produce the amino acid methionine.

Screening for methionine production

For fermentation, a modified basal medium was used (0.3 g K ₂ HPO ₄ ; 0.7 g KH ₂ PO ₄ ; 1.0 g Na ₂ CO ₃ ; 5.0 mg CaCl ₂ ; 0.3 g MgSO ₄ ; 1. 0 mg FeSO ₄ ; 1 L H ₂ O) with sucrose (20.0 g) and NH ₄ Cl (10.0 g) (Chay, BP, Galvez, FCF, and Padolina, WGPULBP (1992). Methionine production by batch fermentation from various carbohydrates. ASEAN Food Journal (Malaysia). The pH value of the medium was 7.2.

Cultivation conditions. Two loops of a 7-day culture of the isolate from Example 8 were inoculated into a 250 ml Erlenmeyer flask containing 30 ml of fermentation medium. Methionine production was analyzed after incubating the flask for 5 days on a rotary shaker (160 rpm) at 30°C. Flasks were prepared in duplicate, and uninoculated flasks served as controls in all experiments.

The presence of methionine in broth cultures of isolates was studied by paper chromatography according to a modification of the method of Khanna and Nag (Khanna et al., "Production of amino acids in vitro tissue culture", Indian Journal of Experimental Biology (1973)). The broth culture was centrifuged at 5000 × g for 20 minutes, and 2 μl of the supernatant was applied 1.5 cm above one edge of an 18 cm × 22 cm Whatman No. 1 filter paper. A 1 μl volume of methionine standard solution (0.1 mg/ ml) was applied along with the supernatant, and the chromatogram was developed in a solvent mixture of n-butanol, acetic acid and water (4:1:1) for 18 hours. The chromatogram was air dried at room temperature, sprayed with 0.15% ninhydrin in butanol and dried again before heating at 60°C for 5 minutes in an oven. The color index value of the ninhydrin-positive spot (bluish-purple) of the supernatant, which corresponds to that of the methionine standard solution, indicates the presence of methionine in the broth culture. The concentration of methionine produced in the broth culture of the isolate was assessed as follows. The ninhydrin-positive spot of the isolate supernatant on the chromatogram was eluted with 10% ethanol, and the spectrophotometric reading of the eluate was recorded at 520 nm. The methionine concentration in the supernatant was determined from the calibration curve. The graph of optical densities as a function of various concentrations (0.1-0.9 mg/ml) of methionine solution served as a calibration curve for methionine.

Isolates that produced methionine were maintained on fresh agar plates, and a stock solution was prepared by suspending two inoculating loops of the organism in an aliquot of a 50% glycerol solution.

Example 10. Introduction of an amino acid-producing bacterial strain into Drosophila melanogaster with a diet to increase their development rate

This example demonstrates the ability to treat Drosophila melanogaster with amino acid-producing bacteria to increase their rate of development when grown on nutrient-deprived diets. This experimental design could be expanded to promote the development of other insects, such as honeybees, that may lack nutrients in their diet.

Treatment regimen

The isolated bacterium Corynebacterium glutamicum, which produces glutamate or methionine, amounted to 10 ⁹ colony forming units (CFU) in a solution mixed with a nutrient substrate for Drosophila flies.

Experimental design

Corynebacterium glutamicum strains that produce glutamate or methionine were grown in nutrient broth at 30°C.

The medium used to feed the flies was a medium containing corn meal, molasses and yeast (11 g/l yeast, 54 g/l yellow corn meal, 5 g/l agar, 66 ml/l molasses and 4.8 ml l propionic acid). All components of this diet, with the exception of propionic acid, were heated together to 80°C in deionized water with constant stirring for 30 minutes and allowed to cool to 60°C. Propionic acid was then mixed in and 50 ml of the diet was aliquoted into separate vials and allowed to cool and solidify. Flies were reared at 26°C, 16:8 h light:dark cycle, and approximately 60% humidity.

To set up an experiment to measure the growth rate of larvae, a diet of a certain composition was used (Piper et al., 2014, Nature Methods). The defined diet eliminated the effects of batch-to-batch variations in the ingredients used for the cornmeal-based diet. In addition, a diet of a certain composition made it possible to exclude individual components to study their effect on the development of flies.

Development rate analysis

On the first day, 100 μl of a suspension of Corynebacterium glutamicum in nutrient broth consisting of 10 ⁹ colony forming units (CFU) was added to the fly feeding medium in five replicates. As a control, nutrient broth without bacteria was added to five more vials of fly feeding medium. Fertilized embryos collected from fruit flies were treated with a 2% hypochlorite solution for five minutes and then washed with sterile water to remove any extracellular microorganisms from the embryos. 10 μl of a suspension of embryos in water (suspension of embryos:water one to three) was added to all bottles containing bacteria inoculated and control food media. The feeding medium for flies with embryos was maintained at a temperature of 26°C, a light:dark cycle of 16:8 hours, and a humidity of about 60% until the end of the experiment. Time to adult emergence and emergence rate were measured in each replicate. From the time the first adult emerged from the pupa, the number of emerging adult flies was counted every 12 h and the rate of emergence was calculated.

Larval mass analysis

To investigate whether the presence of amino acid-producing bacteria can provide an increase in the body weight of developing larvae when reared on a diet of a particular composition, the present inventors obtained larvae that are axenic and monoassociated with a single strain of bacteria. Three different bacteria were used in these assays: Corynebacterium glutamicum, a glutamate-producing strain, Corynebacterium glutamicum, a methionine-producing strain, and E. coli .

First, axenic embryos were obtained. Fertilized embryos were collected from fruit flies over a 6-hour period on grape juice-yeast agar plates. To eliminate any bacterial contamination, embryos were treated with a 2% hypochlorite solution for five minutes and then washed three times with sterile water. One volume of embryos was then suspended in 3 volumes of water.

The defined diet was aliquoted into vials, and nine replicates were used for each condition tested. The conditions were as follows:

1. Feeding medium without added bacteria

2. The feed medium contains a strain of C. glutamicum that produces glutamate (C.glu-Glu)

3. The feed medium contains a strain of C. glutamicum that produces methionine (C.glu-Met)

4. Feeding medium contains E. coli

100 µl of overnight stationary phase cultures were added to each vial of food medium that corresponded to conditions 2, 3 and 4.

To each of nine replicates of each condition, 10 μl of sterile embryo-water suspension was added. The vials were then incubated at 26°C, 60% humidity, 16:8 light:dark cycle.

After 13 days, 10–15 randomly selected larvae from each replicate were sampled and their body areas were measured as an approximate indicator of their biomass and weight. Larvae were scraped from the feeding medium with a sterile spatula, washed in water to clear their bodies of the feeding medium, and each sampled larva was imaged separately for each replicate of each condition. An ImageJ script was used to identify, outline, and measure the body area of the larvae in each image.

When treated with amino acid-producing bacteria, the rate of insect development increases

Embryos that developed on a diet inoculated with an amino acid-producing bacterial strain reached adulthood significantly faster than embryos raised on a sterile diet (Figure 1). In addition, this effect was slightly stronger in female flies than in male flies (Figures 2A and 2B).

When treated with amino acid-producing bacteria, the body weight of larvae increases

Larvae on the diet supplemented with C. glu-Met had the largest body size on average, followed by larvae on the diet supplemented with C. glu-Glu , E. coli , and no bacteria (Fig. 3). This shows that supplementing the insect diet with bacteria that produce the amino acid provided a faster increase in insect biomass than the unsupplemented diet.

Taken together, these data demonstrate that supplementing insect diets with amino acid-producing bacteria produced faster increases in insect biomass than unsupplemented diets. Extending this to other insects such as bees, adding bacteria capable of producing methionine to their diet may provide an increase in their fitness.

Example 11. Insects treated with a solution of purified phages

This example demonstrates the isolation and purification of phages from environmental samples that specifically target specific insect bacteria. This example also demonstrates the effectiveness of the isolated phages against target bacteria in in vitro plaque formation assays, measured by their rate of oxygen consumption and rate of extracellular acidification. Finally, this example demonstrates the effectiveness of phages in vivo, as measured by the ability of the phage to specifically target bacteria in flies by treating them with an isolated phage that counteracts the bacteria. This example demonstrates that a pathogenic bacterium that reduced the fitness of an insect could be eliminated using a phage that specifically targets the bacterium. In particular, Serratia marcescens , which is a pathogenic fly bacterium, can be eliminated using a phage that has been isolated from garden compost.

There are several beneficial and commercially useful insects that are adversely affected by naturally occurring bacterial pathogens. One such example is the bacterium Paenibacillus larvae , which is the cause of foulbrood in honey bees. Using phage to treat foulbrood would go a long way in reducing the huge losses due to foulbrood.

Experimental design

Isolation of specific bacteriophages from natural sources

Bacteriophages that antagonize target bacteria were isolated from environmental source material. Briefly, a saturated culture of Serratia marcescens was diluted in fresh double-strength tryptic soy broth (TSB) and grown for ∼120 min to early log phase at 24–26°C or double-strength Luria-Bertani broth (LB) and grown for ~90 minutes at 37°C. Garden compost was prepared by homogenization in PBS and sterilized by filtration through a 0.2-μm filter. The raw wastewater was sterilized by filtration through a 0.2-μm filter. One volume of the filtered starting material was added to the log phase bacterial cultures and incubation was continued for 24 hours. The enriched starting material was prepared by pelleting the cultures by centrifugation and filtering the supernatant through 0.45 μm pore size membranes.

Phages were isolated by plating samples onto bacterial lawns in a double layer of agar. Stationary bacterial cultures were combined with LB or TSB melted 0.6% agar and poured onto LB or TSB 1.5% agar plates. Once solidified, 2.5 μl of diluted phage samples were spread onto double agar plates and allowed to absorb. The plates were then wrapped and incubated overnight at 25°C (TSA) or 37°C (LB) and assessed for visible plaque formation. Freshly isolated plaques were purified by sequential passage of individual plaques on the target strain by picking up plaques in SM buffer (50 mM Tris-HCl [pH 7.4], 10 mM MgSO4, 100 mM NaCl) and incubating for 15 minutes at 55°C, then repeating the method of applying to a double layer of agar as above using the plaque suspension.

Bacteriophages were successfully isolated from both wastewater and compost, as detailed above. Plaque formation was clearly visible after samples of S. marcescens bacteria were applied to lawns used for enrichment.

Passaging, quantification and propagation of bacteriophages

Propagation and production of phage lysates for use in subsequent experiments were carried out using bacteriophages isolated and purified as described above. Briefly, saturated bacterial cultures were diluted 100-fold in fresh medium and grown for 60–120 min to achieve early log phase growth for efficient phage infection. Phage suspensions or phage lysates were added to early log phase cultures and incubation continued until broth clearing was observed, indicating phage multiplication and bacterial lysis, or until 24 h had passed postinfection. Lysates were collected by pelleting cells by centrifugation at 7197 x g for 20 min. followed by filtering the supernatant through membranes with a pore size of 0.45 or 0.2 μm. Filtered lysates were stored at 4°C.

Enumeration of infectious phage particles was carried out using the double layer agar method. Briefly, serial dilutions of samples were made 1:10 in PBS, and dilutions were spread onto solidified double-layer agar plates prepared with host bacteria as described above. Plaque-forming units (PFU) were counted after overnight incubation to determine the approximate titer of the samples.

In vitro analysis of isolated phages by measuring bacterial respiration

A Seahorse XFe96 analyzer (Agilent) was used to measure the effect of phages on bacteria by monitoring the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) during infection. XFe96 plates were coated the day before experiments with 15 μL of 1 mg/mL poly-L-lysine stock solution per well and dried overnight at 28°C, and XFe96 probes were equilibrated by placing into wells containing 200 μL of XF calibrator. and incubation in the dark at room temperature. The next day, the poly-L-lysine-coated plates were washed twice with ddH2O. Saturated overnight cultures of E. coli BL21 (LB, 37°C) or S. marcescens (TSB, 25°C) were subcultured at 1:100 in the same medium and grown under ventilation for ~2.5 h at 30°C. C. The cultures were then diluted to OD 600 nm ~ 0.02 using the same medium. Treatments were prepared by diluting stock solutions in SM buffer at 10x final concentration and loading 20 μL of 10x solutions into the appropriate sample injection ports of the probe plate. Although the probes were equilibrated in the XFe96 Flux analyzer, bacterial plates were prepared by adding 90 μl of bacterial suspensions or media controls and centrifuged at 3000 rpm. within 10 minutes. After centrifugation, an additional 90 μl of the appropriate medium was carefully added to the wells so as not to disturb bacterial adherence, bringing the total volume to 180 μl per well.

The XFe96 analyzer was run at ~30°C according to mix, wait, read cycles of 1:00, 0:30, 3:00. Four cycles were performed to ensure equilibration/normalization of the bacteria, then 20 μl of treatments were administered and cycles were continued as described above for a total time of approximately 6 hours. Data were analyzed using the Seahorse XFe96 Wave software package.

The effect of the isolated bacteriophages was analyzed by measuring the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of bacteria in a Seahorse XFe96 analyzer. When E. coli was infected with phage T7 and S. marcescens was infected with freshly isolated phage ΦSmVL-C1, significant decreases in OCR were observed after transient peaks in this rate (Fig. 4). For both phages with both host organisms, the Seahorse assay was able to detect successful phage infection without the need for plaque assays. Thus, this method is applicable to detect infection by phages of a host organism that is not amenable to traditional methods for detecting phages.

Transduction assay using SYBR Gold for infection identification

Bacteriophage preparations were prepared for staining by pretreatment with nucleases to remove extraviral nucleic acids that could interfere with the interpretation of the fluorescent signal. Briefly, MgCl2 was added to 10 ml of phage lysate at a final concentration of 10 mM, and both RNase A (Qiagen) and DNase I (Sigma) were added to final concentrations of 10 μg/ml. Samples were incubated for 1 hour at room temperature. After nuclease treatment, 5 ml of lysates were combined with 1 μl of SYBR Gold (Thermo, 10000x) and incubated at room temperature for ~1.5 h. Excess dye was then removed from the samples using Amicon ultrafiltration columns. Briefly, Amicon columns (15 ml, MWCO 10 kDa) were washed by adding 10 ml SM buffer and centrifuged at 5000 x g, 4°C for 5 minutes. Labeled phage samples were then centrifuged in columns at 5000 x g, 4°C until the volume was reduced by approximately 10-fold (15-30 min). To wash the samples, 5 mL of SM buffer was added to each reservoir and centrifugation was repeated, followed by two additional washes. After the third wash, remaining samples were pipetted from Amicon reservoirs and adjusted to approximately 1 mL with SM buffer. To remove larger contaminants, washed and labeled phage samples were centrifuged at 10,000 x g for 2 minutes, and supernatant samples were then filtered through 0.2-μm pore-size membranes into black microtubes and stored at 4°C.

Saturated bacterial cultures ( E. coli MG1655 grown in LB at 37°C, S. marcescens and S. symbiotica grown in TSB at 26°C) were prepared by centrifuging 1 ml aliquots and washing once with 1 ml PBS before final resuspension with 1 ml PBS. Positive control-labeled bacteria were stained by combining 500 μl of washed bacteria and 1 μl of SYBR Gold and incubating for 1 h in the dark at room temperature. Bacteria were pelleted by centrifugation at 8000 x g for 5 min. and washed twice with an equal volume of PBS, followed by resuspension in a final volume of 500 μl PBS. A volume of 25 μl of stained bacteria was combined with 25 μl of SM buffer in a black microtube to which 50 μl of 10% formalin (5% final volume, ~2% formaldehyde) was added and mixed by snapping. Samples were fixed at room temperature for ~3 hours and then washed using Amicon ultrafiltration columns. Briefly, 500 μl of PicoPure water was added to Amicon columns (0.5 ml, MWCO 100 kDa) and centrifuged at 14,000 x g for 5 min. for washing membranes. Fixed samples were diluted by adding 400 μl of PBS and then transferred to prewashed microcentrifuge columns and centrifuged at 14,000 x g for 10 min. The columns were transferred to fresh collection tubes and 500 μl of PBS was added to dilute the fixative remaining in the retentate. Two additional dilutions were then performed with PBS for a total of three washes. The final retentates were diluted to about 100 μl, then the columns were placed upside down in fresh collection tubes and centrifuged at 1000 x g for 2 min. to collect samples. Washed samples were transferred to black microtubes and stored at 4°C.

For transduction experiments and controls, 25 μl of bacteria (or PBS) and 25 μl of SYBR Gold-labeled phage (or SM buffer) were combined in black microtubes and incubated static for 15–20 min. at room temperature to ensure phage adsorption and injection into the receiving bacteria. Immediately after incubation, 50 μl of 10% formalin was added to the samples and fixation was carried out at room temperature for ~4 h. Samples were washed with PBS using Amicon columns as above.

Successful infection of a bacterial host cell by a phage required injection of bacteriophage nucleic acid. Coliphage P1kc, labeled with SYBR Gold and co-incubated with S. marcescens , revealed the presence of fluorescent bacteria by microscopy, validating the use of this assay in phage isolation. As with the Seahorse assay, this approach provided an alternative to traditional phage assays to allow for generalization to organisms not amenable to plaque assays. In addition, the transduction assay using SYBR Gold did not require bacterial growth and was therefore applicable to the analysis of phages targeting difficult-to-culturate or even non-culturable organisms, including endosymbionts such as Buchnera .

Study of the effectiveness of phages in vivo against S. marcescens in Drosophila melanogaster flies

S. marcescens cultures were grown in tryptic soy broth (TSB) at 30°C with constant shaking at 200 rpm.

The medium used to feed the initial fly populations was a medium containing corn meal, molasses and yeast (11 g/L yeast, 54 g/L yellow corn meal, 5 g/L agar, 66 ml/L molasses and 4.8 ml/l propionic acid). All components of this diet, with the exception of propionic acid, were heated together to 80°C in deionized water with constant stirring for 30 minutes and allowed to cool to 60°C. Propionic acid was then mixed in and 50 ml of the diet was aliquoted into separate vials and allowed to cool and solidify. Flies were reared at 26°C, 16:8 h light:dark cycle, and approximately 60% humidity.

To infect flies with S. marcescens, a fine needle (with a tip approximately 10 μm wide) was dipped into a dense overnight culture in the stationary phase, and the chest of the flies was punctured. For this experiment, four replicates of 10 males and 10 females each were infected with S. marcescens using the needle stick method as a positive control for fly mortality. In treatment groups, four replicates of 10 males and 10 females each were inoculated with S. marcescens and a phage solution containing approximately 108 phage particles/ml. Finally, two replicates of 10 males and 10 females each, which were not punctured or otherwise treated, were used as negative controls for mortality.

Flies in all conditions were placed in vials containing food medium and incubated at 26°C, 16:8 h light:dark cycle, 60% humidity. The number of live and dead flies was counted daily for four days after puncture. All flies inoculated with S. marcescens alone were dead 24 hours after treatment. In comparison, more than 60% of the flies in the phage treatment group and all flies in the untreated control group were alive at this time point (Fig. 5). In addition, the majority of flies in the phage treatment group and the negative control group continued to survive for another four days after the experiment was completed.

To determine the cause of fly mortality, dead flies from flies injected with both S. marcescens and S. marcescens +phages were homogenized and plated. Four dead flies from each of four replicates of both S. marcescens and S. marcescens + phage treatments were homogenized in 100 μl of TSB. A 1:100 dilution was also obtained by diluting the homogenate in TSB. 10 μl of the concentrated homogenate, as well as a 1:100 dilution, were plated on TSA plates and incubated overnight at 30°C. When the plates were examined for bacterial growth, all plates containing dead flies that had been inoculated with S. marcescens had a lawn of bacteria growing on them, whereas the plates with dead flies that had been inoculated with S. marcescens + phages had no there were bacteria. This indicated that in the absence of the phage, S. marcescens likely induced septic shock in the flies, leading to their death. However, in the presence of phage, mortality could be due to damage caused by needle puncture.

OTHER IMPLEMENTATION OPTIONS

Although the foregoing invention has been described in sufficient detail by way of illustrations and examples for purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the present invention. All patent and scientific literature disclosures cited herein are expressly incorporated by reference in their entirety.

--->

LIST OF SEQUENCES

<110> Flagship Pioneering, Inc.

<120> COMPOSITIONS AND RELATED METHODS FOR AGRICULTURE

<130> 51215-002WO3

<150>US 62/583,736

<151> 2017-11-09

<150> US 62/450.017

<151> 2017-01-24

<160> 198

<170> PatentIn version 3.5

<210> 1

<211> 1505

<212> DNA

<213> Snodgrassella alvi

<400> 1

gagagtttga tcctggctca gattgaacgc tggcggcatg ccttacacat gcaagtcgaa

60

cggcagcacg gagagcttgc tctctggtgg cgagtggcga acgggtgagt aatgcatcgg

120

aacgtaccga gtaatggggg ataactgtcc gaaaggatgg ctaataccgc atacgccctg

180

agggggaaag cgggggatcg aaagacctcg cgttatttga gcggccgatg ttggattagc

240

tagttggtgg ggtaaaggcc taccaaggcg acgatccata gcgggtctga gaggatgatc

300

cgccacattg ggactgagac acggcccaaa ctcctacggg aggcagcagt ggggaatttt

360

ggacaatggg gggaaccctg atccagccat gccgcgtgtc tgaagaaggc cttcgggttg

420

taaaggactt ttgttaggga agaaaagccg ggtgttaata ccatctggtg ctgacggtac

480

ctaaagaata agcaccggct aactacgtgc cagcagccgc ggtaatacgt agggtgcgag

540

cgttaatcgg aattactggg cgtaaagcga gcgcagacgg ttaattaagt cagatgtgaa

600

atccccgagc tcaacttggg acgtgcattt gaaactggtt aactagagtg tgtcagaggg

660

aggtagaatt ccacgtgtag cagtgaaatg cgtagagatg tggaggaata ccgatggcga

720

aggcagcctc ctgggataac actgacgttc atgctcgaaa gcgtgggtag caaacaggat

780

tagataccct ggtagtccac gccctaaacg atgacaatta gctgttggga cactagatgt

840

cttagtagcg aagctaacgc gtgaaattgt ccgcctgggg agtacggtcg caagattaaa

900

actcaaagga attgacgggg acccgcacaa gcggtggatg atgtggatta attcgatgca

960

acgcgaagaa ccttacctgg tcttgacatg tacggaatct cttagagata ggagagtgcc

1020

ttcgggaacc gtaacacagg tgctgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg

1080

gttaagtccc gcaacgagcg caacccttgt cattagttgc catcattaag ttgggcactc

1140

taatgagact gccggtgaca aaccggagga aggtggggat gacgtcaagt cctcatggcc

1200

cttatgacca gggcttcaca cgtcatacaa tggtcggtac agaggtagc gaagccgcga

1260

ggtgaagcca atctcagaaa gccgatcgta gtccggattg cactctgcaa ctcgagtgca

1320

tgaagtcgga atcgctagta atcgcaggtc agcatactgc ggtgaatacg ttcccgggtc

1380

ttgtacacac cgcccgtcac accatgggag tgggggatac cagaattggg tagactaacc

1440

gcaaggaggt cgcttaacac ggtatgcttc atgactgggg tgaagtcgta acaaggtagc

1500

cgtag

1505

<210> 2

<211> 1541

<212> DNA

<213> Gilliamella apicola

<400> 2

ttaaattgaa gagtttgatc atggctcaga ttgaacgctg gcggcaggct taacacatgc

60

aagtcgaacg gtaacatgag tgcttgcact tgatgacgag tggcggacgg gtgagtaaag

120

tatggggatc tgccgaatgg agggggacaa cagttggaaa cgactgctaa taccgcataa

180

agttgagaga ccaaagcatg ggaccttcgg gccatgcgcc atttgatgaa cccatatggg

240

attagctagt tggtagggta atggcttacc aaggcgacga tctctagctg gtctgagagg

300

atgaccagcc acactggaac tgagacacgg tccagactcc tacgggaggc agcagtgggg

360

aatattgcac aatgggggaa accctgatgc agccatgccg cgtgtatgaa gaaggccttc

420

gggttgtaaa gtactttcgg tgatgaggaa ggtggtgtat ctaataggtg catcaattga

480

cgttaattac agaagaagca ccggctaact ccgtgccagc agccgcggta atacggaggg

540

tgcgagcgtt aatcggaatg actgggcgta aagggcatgt aggcggataa ttaagttagg

600

tgtgaaagcc ctgggctcaa cctaggaatt gcacttaaaa ctggttaact agagtattgt

660

agaggaaggt agaattccac gtgtagcggt gaaatgcgta gagatgtgga ggaataccgg

720

tggcgaaggc ggccttctgg acagatactg acgctgagat gcgaaagcgt ggggagcaaa

780

caggattaga taccctggta gtccacgctg taaacgatgt cgatttggag tttgttgcct

840

agagtgatgg gctccgaagc taacgcgata aatcgaccgc ctggggagta cggccgcaag

900

gttaaaactc aaatgaattg acgggggccc gcacaagcgg tggagcatgt ggtttaattc

960

gatgcaacgc gaagaacctt acctggtctt gacatccaca gaatcttgca gagatgcggg

1020

agtgccttcg ggaactgtga gacaggtgct gcatggctgt cgtcagctcg tgttgtgaaa

1080

tgttgggtta agtcccgcaa cgagcgcaac ccttatcctt tgttgccatc ggttaggccg

1140

ggaactcaaa ggagactgcc gttgataaag cggaggaagg tggggacgac gtcaagtcat

1200

catggccctt acgaccaggg ctacacacgt gctacaatgg cgtatacaaa gggaggcgac

1260

ctcgcgagag caagcggacc tcataaagta cgtctaagtc cggattggag tctgcaactc

1320

gactccatga agtcggaatc gctagtaatc gtgaatcaga atgtcacggt gaatacgttc

1380

ccgggccttg tacacaccgc ccgtcacacc atgggagtgg gttgcaccag aagtagatag

1440

cttaaccttc gggagggcgt ttaccacggt gtggtccatg actggggtga agtcgtaaca

1500

aggtaaccgt aggggaacct gcggttggat cacctcctta c

1541

<210> 3

<211> 1528

<212> DNA

<213> Bartonella apis

<400> 3

aagccaaaat caaattttca acttgagagt ttgatcctgg ctcagaacga acgctggcgg

60

caggcttaac acatgcaagt cgaacgcact tttcggagtg agtggcagac gggtgagtaa

120

cgcgtgggaa tctacctatt tctacggaat aacgcagaga aatttgtgct aataccgtat

180

acgtccttcg ggagaaagat ttatcggaga tagatgagcc cgcgttggat tagctagttg

240

gtgaggtaat ggcccaccaa ggcgacgatc catagctggt ctgagaggat gaccagccac

300

attgggactg agacacggcc cagactccta cgggaggcag cagtggggaa tattggacaa

360

tgggcgcaag cctgatccag ccatgccgcg tgagtgatga aggccctagg gttgtaaagc

420

tctttcaccg gtgaagataa tgacggtaac cggagaagaa gccccggcta acttcgtgcc

480

agcagccgcg gtaatacgaa gggggctagc gttgttcgga tttactgggc gtaaagcgca

540

cgtaggcgga tatttaagtc aggggtgaaa tcccggggct caaccccgga actgcctttg

600

atactggata tcttgagtat ggaagaggta agtggaattc cgagtgtaga ggtgaaattc

660

gtagatattc ggaggaacac cagtggcgaa ggcggcttac tggtccatta ctgacgctga

720

ggtgcgaaag cgtggggagc aaacaggatt agataccctg gtagtccacg ctgtaaacga

780

tgaatgttag ccgttggaca gtttactgtt cggtggcgca gctaacgcat taaacattcc

840

gcctggggag tacggtcgca agattaaaac tcaaaggaat tgacgggggc ccgcacaagc

900

ggtggagcat gtggtttaat tcgaagcaac gcgcagaacc ttaccagccc ttgacatccc

960

gatcgcggat ggtggagaca ccgtctttca gttcggctgg atcggtgaca ggtgctgcat

1020

ggctgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctc

1080

gcccttagtt gccatcattt agttgggcac tctaagggga ctgccggtga taagccgaga

1140

ggaaggtggg gatgacgtca agtcctcatg gcccttacgg gctgggctac acacgtgcta

1200

caatggtggt gacagtgggc agcgagaccg cgaggtcgag ctaatctcca aaagccatct

1260

cagttcggat tgcactctgc aactcgagtg catgaagttg gaatcgctag taatcgtgga

1320

tcagcatgcc acggtgaata cgttcccggg ccttgtacac accgcccgtc acaccatggg

1380

agttggtttt acccgaaggt gctgtgctaa ccgcaaggag gcaggcaacc acggtagggt

1440

cagcgactgg ggtgaagtcg taacaaggta gccgtagggg aacctgcggc tggatcacct

1500

cctttctaag gaagatgaag aattggaa

1528

<210> 4

<211> 1390

<212> DNA

<213> Parasaccharibacter apium

<220>

<221> other_sign

<222> (643)..(756)

<223> n represents a, g, c or t

<400> 4

ctaccatgca agtcgcacga aacctttcgg ggttagtggc ggacgggtga gtaacgcgtt

60

aggaacctat ctggaggtgg gggataacat cgggaaactg gtgctaatac cgcatgatgc

120

ctgaggggcca aagggagat ccgccattgg aggggcctgc gttcgattag ctagttggtt

180

gggtaaaggc tgaccaaggc gatgatcgat agctggtttg agaggatgat cagccacact

240

gggactgaga cacggcccag actcctacgg gaggcagcag tggggaatat tggacaatgg

300

gggcaaccct gatccagcaa tgccgcgtgt gtgaagaagg tcttcggatt gtaaagcact

360

ttcactaggg aagatgatga cggtacctag agaagaagcc ccggctaact tcgtgccagc

420

agccgcggta atacgaaggg ggctagcgtt gctcggaatg actgggcgta aagggcgcgt

480

aggctgtttg tacagtcaga tgtgaaatcc ccgggcttaa cctgggaact gcatttgata

540

cgtgcagact agagtccgag agagggttgt ggaattccca gtgtagaggt gaaattcgta

600

gatattggga agaacaccgg ttgcgaaggc ggcaacctgg ctnnnnnnnn nnnnnnnnnn

660

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn

720

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnngagc taacgcgtta agcacaccgc

780

ctggggagta cggccgcaag gttgaaactc aaaggaattg acgggggccc gcacaagcgg

840

tggagcatgt ggtttaattc gaagcaacgc gcagaacctt accagggctt gcatggggag

900

gctgtattca gagatggata tttcttcgga cctcccgcac aggtgctgca tggctgtcgt

960

cagctcgtgt cgtgagatgt tgggttaagt cccgcaacga gcgcaaccct tgtctttagt

1020

tgccatcacg tctgggtggg cactctagag agactgccgg tgacaagccg gaggaaggtg

1080

gggatgacgt caagtcctca tggcccttat gtcctgggct acaacgtgc tacaatggcg

1140

gtgacagagg gatgctacat ggtgacatgg tgctgatctc aaaaaaccgt ctcagttcgg

1200

attgtactct gcaactcgag tgcatgaagg tggaatcgct agtaatcgcg gatcagcatg

1260

ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg tcacaccatg ggagttggtt

1320

tgaccttaag ccggtgagcg aaccgcaagg aacgcagccg accaccggtt cgggttcagc

1380

gactggggga

1390

<210> 5

<211> 1549

<212> DNA

<213> Enterococcus

<400> 5

aggtgatcca gccgcacctt ccgatacggc taccttgtta cgacttcacc ccaatcatct

60

atcccacctt aggcggctgg ctccaaaaag gttacctcac cgacttcggg tgttacaaac

120

tctcgtggtg tgacgggcgg tgtgtacaag gcccgggaac gtattcaccg cggcgtgctg

180

atccgcgatt actagcgatt ccggcttcat gcaggcgagt tgcagcctgc aatccgaact

240

gagagaagct ttaagagatt tgcatgacct cgcggtctag cgactcgttg tacttcccat

300

tgtagcacgt gtgtagccca ggtcataagg ggcatgatga tttgacgtca tccccacctt

360

cctccggttt gtcaccggca gtctcgctag agtgcccaac taaatgatgg caactaacaa

420

taagggttgc gctcgttgcg ggacttaacc caacatctca cgacacgagc tgacgacaac

480

catgcaccac ctgtcacttt gtccccgaag ggaaagctct atctctagag tggtcaaagg

540

atgtcaagac ctggtaaggt tcttcgcgtt gcttcgaatt aaaccacatg ctccaccgct

600

tgtgcgggcc cccgtcaatt cctttgagtt tcaaccttgc ggtcgtactc cccaggcgga

660

gtgcttaatg cgtttgctgc agcactgaag ggcggaaacc ctccaacact tagcactcat

720

cgtttacggc gtggactacc agggtatcta atcctgtttg ctccccacgc tttcgagcct

780

cagcgtcagt tacagaccag agagccgcct tcgccactgg tgttcctcca tatatctacg

840

catttcaccg ctacacatgg aattccactc tcctcttctg cactcaagtc tcccagtttc

900

caatgaccct ccccggttga gccggggggct ttcacatcag acttaagaaa ccgcctgcgc

960

tcgctttacg cccaataaat ccggacaacg cttgccacct acgtattacc gcggctgctg

1020

gcacgtagtt agccgtggct ttctggttag ataccgtcag gggacgttca gttactaacg

1080

tccttgttct tctctaacaa cagagtttta cgatccgaaa accttcttca ctcacgcggc

1140

gttgctcggt cagactttcg tccattgccg aagattccct actgctgcct cccgtaggag

1200

tctgggccgt gtctcagtcc cagtgtggcc gatcaccctc tcaggtcggc tatgcatcgt

1260

ggccttggtg agccgttacc tcaccaacta gctaatgcac cgcgggtcca tccatcagcg

1320

acacccgaaa gcgcctttca ctcttatgcc atgcggcata aactgttatg cggtattagc

1380

acctgtttcc aagtgttatc cccctctgat gggtaggtta cccacgtgtt actcacccgt

1440

ccgccactcc tctttccaat tgagtgcaag cactcgggag gaaagaagcg ttcgacttgc

1500

atgtattagg cacgccgcca gcgttcgtcc tgagccagga tcaaactct

1549

<210> 6

<211> 1541

<212> DNA

<213> Delftia

<400> 6

cagaaaggag gtgatccagc cgcaccttcc gatacggcta ccttgttacg acttcacccc

60

agtcacgaac cccgccgtgg taagcgccct ccttgcggtt aggctaccta cttctggcga

120

gacccgctcc catggtgtga cgggcggtgt gtacaagacc cgggaacgta ttcaccgcgg

180

catgctgatc cgcgattact agcgattccg acttcacgca gtcgagttgc agactgcgat

240

ccggactacg actggtttta tgggattagc tccccctcgc gggttggcaa ccctctgtac

300

cagccattgt atgacgtgtg tagccccacc tataagggcc atgaggactt gacgtcatcc

360

ccaccttcct ccggtttgtc accggcagtc tcattagagt gctcaactga atgtagcaac

420

taatgacaag ggttgcgctc gttgcgggac ttaacccaac atctcacgac acgagctgac

480

gacagccatg cagcacctgt gtgcaggttc tctttcgagc acgaatccat ctctggaaac

540

ttcctgccat gtcaaaggtg ggtaaggttt ttcgcgttgc atcgaattaa accacatcat

600

ccaccgcttg tgcgggtccc cgtcaattcc tttgagtttc aaccttgcgg ccgtactccc

660

caggcggtca acttcacgcg ttagcttcgt tactgagaaa actaattccc aacaaccagt

720

tgacatcgtt tagggcgtgg actaccaggg tatctaatcc tgtttgctcc ccacgctttc

780

gtgcatgagc gtcagtacag gtccagggga ttgccttcgc catcggtgtt cctccgcata

840

tctacgcatt tcactgctac acgcggaatt ccatccccct ctaccgtact ctagccatgc

900

agtcacaaat gcagttccca ggttgagccc ggggatttca catctgtctt acataaccgc

960

ctgcgcacgc tttacgccca gtaattccga ttaacgctcg caccctacgt attaccgcgg

1020

ctgctggcac gtagttagcc ggtgcttatt cttacggtac cgtcatgggc cccctgtatt

1080

agaaggagct ttttcgttcc gtacaaaagc agtttacaac ccgaaggcct tcatcctgca

1140

cgcggcattg ctggatcagg ctttcgccca ttgtccaaaa ttccccactg ctgcctcccg

1200

taggagtctg ggccgtgtct cagtcccagt gtggctggtc gtcctctcag accagctaca

1260

gatcgtcggc ttggtaagct tttatcccac caactaccta atctgccatc ggccgctcca

1320

atcgcgcgag gcccgaaggg cccccgcttt catcctcaga tcgtatgcgg tattagctac

1380

tctttcgagt agttatcccc cacgactggg cacgttccga tgtattactc acccgttcgc

1440

cactcgtcag cgtccgaaga cctgttaccg ttcgacttgc atgtgtaagg catgccgcca

1500

gcgttcaatc tgagccagga tcaaactcta cagttcgatc t

1541

<210> 7

<211> 1502

<212> DNA

<213> Pelomonas

<220>

<221> other_sign

<222> (192)..(193)

<223> n represents a, c, g or t

<220>

<221> other_sign

<222> (832)..(833)

<223> n represents a, c, g or t

<400> 7

atcctggctc agattgaacg ctggcggcat gccttacaca tgcaagtcga acggtaacag

60

gttaagctga cgagtggcga acgggtgagt aatatatcgg aacgtgccca gtcgtggggg

120

ataactgctc gaaagagcag ctaataccgc atacgacctg agggtgaaag cgggggatcg

180

caagacctcg cnngattgga gcggccgata tcagattagg tagttggtgg ggtaaaggcc

240

caccaagcca acgatctgta gctggtctga gaggacgacc agccacactg ggactgagac

300

acggcccaga ctcctacggg aggcagcagt ggggaatttt ggacaatggg cgcaagcctg

360

atccagccat gccgcgtgcg ggaagaaggc cttcgggttg taaaccgctt ttgtcaggga

420

agaaaaggtt ctggttaata cctgggactc atgacggtac ctgaagaata agcaccggct

480

aactacgtgc cagcagccgc ggtaatacgt agggtgcaag cgttaatcgg aattactggg

540

cgtaaagcgt gcgcaggcgg ttatgcaaga cagaggtgaa atccccgggc tcaacctggg

600

aactgccttt gtgactgcat agctagagta cggtagaggg ggatggaatt ccgcgtgtag

660

cagtgaaatg cgtagatatg cggaggaaca ccgatggcga aggcaatccc ctggacctgt

720

actgacgctc atgcacgaaa gcgtggggag caaacaggat tagataccct ggtagtccac

780

gccctaaacg atgtcaactg gttgttggga gggtttcttc tcagtaacgt anntaacgcg

840

tgaagttgac cgcctgggga gtacggccgc aaggttgaaa ctcaaaggaa ttgacgggga

900

cccgcacaag cggtggatga tgtggtttaa ttcgatgcaa cgcgaaaaac cttacctacc

960

cttgacatgc caggaatcct gaagagattt gggagtgctc gaaagagaac ctggacacag

1020

gtgctgcatg gccgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc

1080

gcaacccttg tcattagttg ctacgaaagg gcactctaat gagactgccg gtgacaaacc

1140

gggaggaaggt ggggatgacg tcaggtcatc atggccctta tgggtagggc tacacacgtc

1200

atacaatggc cgggacagag ggctgccaac ccgcgagggg gagctaatcc cagaaacccg

1260

gtcgtagtcc ggatcgtagt ctgcaactcg actgcgtgaa gtcggaatcg ctagtaatcg

1320

cggatcagct tgccgcggtg aatacgttcc cgggtcttgt acacaccgcc cgtcacacca

1380

tgggagcggg ttctgccaga agtagttagc ctaaccgcaa ggagggcgat taccacggca

1440

gggttcgtga ctggggtgaa gtcgtaacaa ggtagccgta tcggaaggtg cggctggatc

1500

ac

1502

<210> 8

<211> 34

<212> PROTEIN

<213> Lactococcus lactis

<400> 8

Ile Thr Ser Ile Ser Leu Cys Thr Pro Gly Cys Lys Thr Gly Ala Leu

1 5 10 15

Met Gly Cys Asn Met Lys Thr Ala Thr Cys His Cys Ser Ile His Val

20 25 30

Ser Lys

<210> 9

<211> 22

<212> PROTEIN

<213> Staphylococcus epidermis

<400> 9

Ile Ala Ser Lys Phe Ile Cys Thr Pro Gly Cys Ala Lys Thr Gly Ser

1 5 10 15

Phe Asn Ser Tyr Cys Cys

20

<210> 10

<211> 44

<212> PROTEIN

<213> Pediococcus acidilactici

<400> 10

Lys Tyr Tyr Gly Asn Gly Val Thr Cys Gly Lys His Ser Cys Ser Val

1 5 10 15

Asp Trp Gly Lys Ala Thr Thr Cys Ile Ile Asn Asn Gly Ala Met Ala

20 25 30

Trp Ala Thr Gly Gly His Gln Gly Asn His Lys Cys

35 40

<210> 11

<211> 44

<212> PROTEIN

<213> Enterococcus faecium

<400> 11

Ala Thr Arg Ser Tyr Gly Asn Gly Val Tyr Cys Asn Asn Ser Lys Cys

1 5 10 15

Trp Val Asn Trp Gly Glu Ala Lys Glu Asn Ile Ala Gly Ile Val Ile

20 25 30

Ser Gly Trp Ala Ser Gly Leu Ala Gly Met Gly His

35 40

<210> 12

<211> 39

<212> PROTEIN

<213> Streptococcus lactis

<400> 12

Gly Thr Trp Asp Asp Ile Gly Gln Gly Ile Gly Arg Val Ala Tyr Trp

1 5 10 15

Val Gly Lys Ala Met Gly Asn Met Ser Asp Val Asn Gln Ala Ser Arg

20 25 30

Ile Asn Arg Lys Lys Lys His

35

<210> 13

<211> 48

<212> PROTEIN

<213> Lactobacillus johnsonii

<400> 13

Asn Arg Trp Gly Asp Thr Val Leu Ser Ala Ala Ser Gly Ala Gly Thr

1 5 10 15

Gly Ile Lys Ala Cys Lys Ser Phe Gly Pro Trp Gly Met Ala Ile Cys

20 25 30

Gly Val Gly Gly Ala Ala Ile Gly Gly Tyr Phe Gly Tyr Thr His Asn

35 40 45

<210> 14

<211> 70

<212> PROTEIN

<213> Enterococcus faecalis

<400> 14

Met Ala Lys Glu Phe Gly Ile Pro Ala Ala Val Ala Gly Thr Val Leu

1 5 10 15

Asn Val Val Glu Ala Gly Gly Trp Val Thr Thr Ile Val Ser Ile Leu

20 25 30

Thr Ala Val Gly Ser Gly Gly Leu Ser Leu Leu Ala Ala Ala Gly Arg

35 40 45

Glu Ser Ile Lys Ala Tyr Leu Lys Lys Glu Ile Lys Lys Lys Gly Lys

50 55 60

Arg Ala Val Ile Ala Trp

65 70

<210> 15

<211> 51

<212> PROTEIN

<213> Staphylococcus aureus

<400> 15

Met Ser Trp Leu Asn Phe Leu Lys Tyr Ile Ala Lys Tyr Gly Lys Lys

1 5 10 15

Ala Val Ser Ala Ala Trp Lys Tyr Lys Gly Lys Val Leu Glu Trp Leu

20 25 30

Asn Val Gly Pro Thr Leu Glu Trp Val Trp Gln Lys Leu Lys Lys Ile

35 40 45

Ala Gly Leu

50

<210> 16

<211> 43

<212> PROTEIN

<213> Lactococcus garvieae

<400> 16

Ile Gly Gly Ala Leu Gly Asn Ala Leu Asn Gly Leu Gly Thr Trp Ala

1 5 10 15

Asn Met Met Asn Gly Gly Gly Phe Val Asn Gln Trp Gln Val Tyr Ala

20 25 30

Asn Lys Gly Lys Ile Asn Gln Tyr Arg Pro Tyr

35 40

<210> 17

<211> 103

<212> PROTEIN

<213> Escherichia coli

<400> 17

Met Arg Thr Leu Thr Leu Asn Glu Leu Asp Ser Val Ser Gly Gly Ala

1 5 10 15

Ser Gly Arg Asp Ile Ala Met Ala Ile Gly Thr Leu Ser Gly Gln Phe

20 25 30

Val Ala Gly Gly Ile Gly Ala Ala Ala Gly Gly Val Ala Gly Gly Ala

35 40 45

Ile Tyr Asp Tyr Ala Ser Thr His Lys Pro Asn Pro Ala Met Ser Pro

50 55 60

Ser Gly Leu Gly Gly Thr Ile Lys Gln Lys Pro Glu Gly Ile Pro Ser

65 70 75 80

Glu Ala Trp Asn Tyr Ala Ala Gly Arg Leu Cys Asn Trp Ser Pro Asn

85 90 95

Asn Leu Ser Asp Val Cys Leu

100

<210> 18

<211> 339

<212> PROTEIN

<213> Cp1

<400> 18

Met Val Lys Lys Asn Asp Leu Phe Val Asp Val Ser Ser His Asn Gly

1 5 10 15

Tyr Asp Ile Thr Gly Ile Leu Glu Gln Met Gly Thr Thr Asn Thr Ile

20 25 30

Ile Lys Ile Ser Glu Ser Thr Thr Tyr Leu Asn Pro Cys Leu Ser Ala

35 40 45

Gln Val Glu Gln Ser Asn Pro Ile Gly Phe Tyr His Phe Ala Arg Phe

50 55 60

Gly Gly Asp Val Ala Glu Ala Glu Arg Glu Ala Gln Phe Phe Leu Asp

65 70 75 80

Asn Val Pro Met Gln Val Lys Tyr Leu Val Leu Asp Tyr Glu Asp Asp

85 90 95

Pro Ser Gly Asp Ala Gln Ala Asn Thr Asn Ala Cys Leu Arg Phe Met

100 105 110

Gln Met Ile Ala Asp Ala Gly Tyr Lys Pro Ile Tyr Tyr Ser Tyr Lys

115 120 125

Pro Phe Thr His Asp Asn Val Asp Tyr Gln Gln Ile Leu Ala Gln Phe

130 135 140

Pro Asn Ser Leu Trp Ile Ala Gly Tyr Gly Leu Asn Asp Gly Thr Ala

145 150 155 160

Asn Phe Glu Tyr Phe Pro Ser Met Asp Gly Ile Arg Trp Trp Gln Tyr

165 170 175

Ser Ser Asn Pro Phe Asp Lys Asn Ile Val Leu Leu Asp Asp Glu Glu

180 185 190

Asp Asp Lys Pro Lys Thr Ala Gly Thr Trp Lys Gln Asp Ser Lys Gly

195 200 205

Trp Trp Phe Arg Arg Asn Asn Gly Ser Phe Pro Tyr Asn Lys Trp Glu

210 215 220

Lys Ile Gly Gly Val Trp Tyr Tyr Phe Asp Ser Lys Gly Tyr Cys Leu

225 230 235 240

Thr Ser Glu Trp Leu Lys Asp Asn Glu Lys Trp Tyr Tyr Leu Lys Asp

245 250 255

Asn Gly Ala Met Ala Thr Gly Trp Val Leu Val Gly Ser Glu Trp Tyr

260 265 270

Tyr Met Asp Asp Ser Gly Ala Met Val Thr Gly Trp Val Lys Tyr Lys

275 280 285

Asn Asn Trp Tyr Tyr Met Thr Asn Glu Arg Gly Asn Met Val Ser Asn

290 295 300

Glu Phe Ile Lys Ser Gly Lys Gly Trp Tyr Phe Met Asn Thr Asn Gly

305 310 315 320

Glu Leu Ala Asp Asn Pro Ser Phe Thr Lys Glu Pro Asp Gly Leu Ile

325 330 335

Thr Val Ala

<210> 19

<211> 296

<212> PROTEIN

<213> Dp-1

<400> 19

Met Gly Val Asp Ile Glu Lys Gly Val Ala Trp Met Gln Ala Arg Lys

1 5 10 15

Gly Arg Val Ser Tyr Ser Met Asp Phe Arg Asp Gly Pro Asp Ser Tyr

20 25 30

Asp Cys Ser Ser Ser Met Tyr Tyr Ala Leu Arg Ser Ala Gly Ala Ser

35 40 45

Ser Ala Gly Trp Ala Val Asn Thr Glu Tyr Met His Ala Trp Leu Ile

50 55 60

Glu Asn Gly Tyr Glu Leu Ile Ser Glu Asn Ala Pro Trp Asp Ala Lys

65 70 75 80

Arg Gly Asp Ile Phe Ile Trp Gly Arg Lys Gly Ala Ser Ala Gly Ala

85 90 95

Gly Gly His Thr Gly Met Phe Ile Asp Ser Asp Asn Ile Ile His Cys

100 105 110

Asn Tyr Ala Tyr Asp Gly Ile Ser Val Asn Asp His Asp Glu Arg Trp

115 120 125

Tyr Tyr Ala Gly Gln Pro Tyr Tyr Tyr Val Tyr Arg Leu Thr Asn Ala

130 135 140

Asn Ala Gln Pro Ala Glu Lys Lys Leu Gly Trp Gln Lys Asp Ala Thr

145 150 155 160

Gly Phe Trp Tyr Ala Arg Ala Asn Gly Thr Tyr Pro Lys Asp Glu Phe

165 170 175

Glu Tyr Ile Glu Glu Asn Lys Ser Trp Phe Tyr Phe Asp Asp Gln Gly

180 185 190

Tyr Met Leu Ala Glu Lys Trp Leu Lys His Thr Asp Gly Asn Trp Tyr

195 200 205

Trp Phe Asp Arg Asp Gly Tyr Met Ala Thr Ser Trp Lys Arg Ile Gly

210 215 220

Glu Ser Trp Tyr Tyr Phe Asn Arg Asp Gly Ser Met Val Thr Gly Trp

225 230 235 240

Ile Lys Tyr Tyr Asp Asn Trp Tyr Tyr Cys Asp Ala Thr Asn Gly Asp

245 250 255

Met Lys Ser Asn Ala Phe Ile Arg Tyr Asn Asp Gly Trp Tyr Leu Leu

260 265 270

Leu Pro Asp Gly Arg Leu Ala Asp Lys Pro Gln Phe Thr Val Glu Pro

275 280 285

Asp Gly Leu Ile Thr Ala Lys Val

290 295

<210> 20

<211> 233

<212> PROTEIN

<213> gamma

<400> 20

Met Glu Ile Gln Lys Lys Leu Val Asp Pro Ser Lys Tyr Gly Thr Lys

1 5 10 15

Cys Pro Tyr Thr Met Lys Pro Lys Tyr Ile Thr Val His Asn Thr Tyr

20 25 30

Asn Asp Ala Pro Ala Glu Asn Glu Val Ser Tyr Met Ile Ser Asn Asn

35 40 45

Asn Glu Val Ser Phe His Ile Ala Val Asp Asp Lys Lys Ala Ile Gln

50 55 60

Gly Ile Pro Leu Glu Arg Asn Ala Trp Ala Cys Gly Asp Gly Asn Gly

65 70 75 80

Ser Gly Asn Arg Gln Ser Ile Ser Val Glu Ile Cys Tyr Ser Lys Ser

85 90 95

Gly Gly Asp Arg Tyr Tyr Lys Ala Glu Asp Asn Ala Val Asp Val Val

100 105 110

Arg Gln Leu Met Ser Met Tyr Asn Ile Pro Ile Glu Asn Val Arg Thr

115 120 125

His Gln Ser Trp Ser Gly Lys Tyr Cys Pro His Arg Met Leu Ala Glu

130 135 140

Gly Arg Trp Gly Ala Phe Ile Gln Lys Val Lys Asn Gly Asn Val Ala

145 150 155 160

Thr Thr Ser Pro Thr Lys Gln Asn Ile Ile Gln Ser Gly Ala Phe Ser

165 170 175

Pro Tyr Glu Thr Pro Asp Val Met Gly Ala Leu Thr Ser Leu Lys Met

180 185 190

Thr Ala Asp Phe Ile Leu Gln Ser Asp Gly Leu Thr Tyr Phe Ile Ser

195 200 205

Lys Pro Thr Ser Asp Ala Gln Leu Lys Ala Met Lys Glu Tyr Leu Asp

210 215 220

Arg Lys Gly Trp Trp Tyr Glu Val Lys

225 230

<210> 21

<211> 481

<212> PROTEIN

<213>MR11

<400> 21

Met Gln Ala Lys Leu Thr Lys Lys Glu Phe Ile Glu Trp Leu Lys Thr

1 5 10 15

Ser Glu Gly Lys Gln Phe Asn Val Asp Leu Trp Tyr Gly Phe Gln Cys

20 25 30

Phe Asp Tyr Ala Asn Ala Gly Trp Lys Val Leu Phe Gly Leu Leu Leu

35 40 45

Lys Gly Leu Gly Ala Lys Asp Ile Pro Phe Ala Asn Asn Phe Asp Gly

50 55 60

Leu Ala Thr Val Tyr Gln Asn Thr Pro Asp Phe Leu Ala Gln Pro Gly

65 70 75 80

Asp Met Val Val Phe Gly Ser Asn Tyr Gly Ala Gly Tyr Gly His Val

85 90 95

Ala Trp Val Ile Glu Ala Thr Leu Asp Tyr Ile Ile Val Tyr Glu Gln

100 105 110

Asn Trp Leu Gly Gly Gly Trp Thr Asp Arg Ile Glu Gln Pro Gly Trp

115 120 125

Gly Trp Glu Lys Val Thr Arg Arg Gln His Ala Tyr Asp Phe Pro Met

130 135 140

Trp Phe Ile Arg Pro Asn Phe Lys Ser Glu Thr Ala Pro Arg Ser Ile

145 150 155 160

Gln Ser Pro Thr Gln Ala Ser Lys Lys Glu Thr Ala Lys Pro Gln Pro

165 170 175

Lys Ala Val Glu Leu Lys Ile Ile Lys Asp Val Val Lys Gly Tyr Asp

180 185 190

Leu Pro Lys Arg Gly Gly Asn Pro Lys Gly Ile Val Ile His Asn Asp

195 200 205

Ala Gly Ser Lys Gly Ala Thr Ala Glu Ala Tyr Arg Asn Gly Leu Val

210 215 220

Asn Ala Pro Leu Ser Arg Leu Glu Ala Gly Ile Ala His Ser Tyr Val

225 230 235 240

Ser Gly Asn Thr Val Trp Gln Ala Leu Asp Glu Ser Gln Val Gly Trp

245 250 255

His Thr Ala Asn Gln Leu Gly Asn Lys Tyr Tyr Tyr Gly Ile Glu Val

260 265 270

Cys Gln Ser Met Gly Ala Asp Asn Ala Thr Phe Leu Lys Asn Glu Gln

275 280 285

Ala Thr Phe Gln Glu Cys Ala Arg Leu Leu Lys Lys Trp Gly Leu Pro

290 295 300

Ala Asn Arg Asn Thr Ile Arg Leu His Asn Glu Phe Thr Ser Thr Ser

305 310 315 320

Cys Pro His Arg Ser Ser Val Leu His Thr Gly Phe Asp Pro Val Thr

325 330 335

Arg Gly Leu Leu Pro Glu Asp Lys Gln Leu Gln Leu Lys Asp Tyr Phe

340 345 350

Ile Lys Gln Ile Arg Val Tyr Met Asp Gly Lys Ile Pro Val Ala Thr

355 360 365

Val Ser Asn Glu Ser Ser Ala Ser Ser Asn Thr Val Lys Pro Val Ala

370 375 380

Ser Ala Trp Lys Arg Asn Lys Tyr Gly Thr Tyr Tyr Met Glu Glu Asn

385 390 395 400

Ala Arg Phe Thr Asn Gly Asn Gln Pro Ile Thr Val Arg Lys Ile Gly

405 410 415

Pro Phe Leu Ser Cys Pro Val Ala Tyr Gln Phe Gln Pro Gly Gly Tyr

420 425 430

Cys Asp Tyr Thr Glu Val Met Leu Gln Asp Gly His Val Trp Val Gly

435 440 445

Tyr Thr Trp Glu Gly Gln Arg Tyr Tyr Leu Pro Ile Arg Thr Trp Asn

450 455 460

Gly Ser Ala Pro Pro Asn Gln Ile Leu Gly Asp Leu Trp Gly Glu Ile

465 470 475 480

Ser

<210> 22

<211> 239

<212> PROTEIN

<213>B30

<400> 22

Met Val Ile Asn Ile Glu Gln Ala Ile Ala Trp Met Ala Ser Arg Lys

1 5 10 15

Gly Lys Val Thr Tyr Ser Met Asp Tyr Arg Asn Gly Pro Ser Ser Tyr

20 25 30

Asp Cys Ser Ser Ser Val Tyr Phe Ala Leu Arg Ser Ala Gly Ala Ser

35 40 45

Asp Asn Gly Trp Ala Val Asn Thr Glu Tyr Glu His Asp Trp Leu Ile

50 55 60

Lys Asn Gly Tyr Val Leu Ile Ala Glu Asn Thr Asn Trp Asn Ala Gln

65 70 75 80

Arg Gly Asp Ile Phe Ile Trp Gly Lys Arg Gly Ala Ser Ala Gly Ala

85 90 95

Phe Gly His Thr Gly Met Phe Val Asp Pro Asp Asn Ile Ile His Cys

100 105 110

Asn Tyr Gly Tyr Asn Ser Ile Thr Val Asn Asn His Asp Glu Ile Trp

115 120 125

Gly Tyr Asn Gly Gln Pro Tyr Val Tyr Ala Tyr Arg Tyr Ser Gly Lys

130 135 140

Gln Ser Asn Ala Lys Val Asp Asn Lys Ser Val Val Ser Lys Phe Glu

145 150 155 160

Lys Glu Leu Asp Val Asn Thr Pro Leu Ser Asn Ser Asn Met Pro Tyr

165 170 175

Tyr Glu Ala Thr Ile Ser Glu Asp Tyr Tyr Val Glu Ser Lys Pro Asp

180 185 190

Val Asn Ser Thr Asp Lys Glu Leu Leu Val Ala Gly Thr Arg Val Arg

195 200 205

Val Tyr Glu Lys Val Lys Gly Trp Ala Arg Ile Gly Ala Pro Gln Ser

210 215 220

Asn Gln Trp Val Glu Asp Ala Tyr Leu Ile Asp Ala Thr Asp Met

225 230 235

<210> 23

<211> 495

<212> PROTEIN

<213>K

<400> 23

Met Ala Lys Thr Gln Ala Glu Ile Asn Lys Arg Leu Asp Ala Tyr Ala

1 5 10 15

Lys Gly Thr Val Asp Ser Pro Tyr Arg Val Lys Lys Ala Thr Ser Tyr

20 25 30

Asp Pro Ser Phe Gly Val Met Glu Ala Gly Ala Ile Asp Ala Asp Gly

35 40 45

Tyr Tyr His Ala Gln Cys Gln Asp Leu Ile Thr Asp Tyr Val Leu Trp

50 55 60

Leu Thr Asp Asn Lys Val Arg Thr Trp Gly Asn Ala Lys Asp Gln Ile

65 70 75 80

Lys Gln Ser Tyr Gly Thr Gly Phe Lys Ile His Glu Asn Lys Pro Ser

85 90 95

Thr Val Pro Lys Lys Gly Trp Ile Ala Val Phe Thr Ser Gly Ser Tyr

100 105 110

Glu Gln Trp Gly His Ile Gly Ile Val Tyr Asp Gly Gly Asn Thr Ser

115 120 125

Thr Phe Thr Ile Leu Glu Gln Asn Trp Asn Gly Tyr Ala Asn Lys Lys

130 135 140

Pro Thr Lys Arg Val Asp Asn Tyr Tyr Gly Leu Thr His Phe Ile Glu

145 150 155 160

Ile Pro Val Lys Ala Gly Thr Thr Val Lys Lys Glu Thr Ala Lys Lys

165 170 175

Ser Ala Ser Lys Thr Pro Ala Pro Lys Lys Lys Ala Thr Leu Lys Val

180 185 190

Ser Lys Asn His Ile Asn Tyr Thr Met Asp Lys Arg Gly Lys Lys Pro

195 200 205

Glu Gly Met Val Ile His Asn Asp Ala Gly Arg Ser Ser Gly Gln Gln

210 215 220

Tyr Glu Asn Ser Leu Ala Asn Ala Gly Tyr Ala Arg Tyr Ala Asn Gly

225 230 235 240

Ile Ala His Tyr Tyr Gly Ser Glu Gly Tyr Val Trp Glu Ala Ile Asp

245 250 255

Ala Lys Asn Gln Ile Ala Trp His Thr Gly Asp Gly Thr Gly Ala Asn

260 265 270

Ser Gly Asn Phe Arg Phe Ala Gly Ile Glu Val Cys Gln Ser Met Ser

275 280 285

Ala Ser Asp Ala Gln Phe Leu Lys Asn Glu Gln Ala Val Phe Gln Phe

290 295 300

Thr Ala Glu Lys Phe Lys Glu Trp Gly Leu Thr Pro Asn Arg Lys Thr

305 310 315 320

Val Arg Leu His Met Glu Phe Val Pro Thr Ala Cys Pro His Arg Ser

325 330 335

Met Val Leu His Thr Gly Phe Asn Pro Val Thr Gln Gly Arg Pro Ser

340 345 350

Gln Ala Ile Met Asn Lys Leu Lys Asp Tyr Phe Ile Lys Gln Ile Lys

355 360 365

Asn Tyr Met Asp Lys Gly Thr Ser Ser Ser Thr Val Val Lys Asp Gly

370 375 380

Lys Thr Ser Ser Ala Ser Thr Pro Ala Thr Arg Pro Val Thr Gly Ser

385 390 395 400

Trp Lys Lys Asn Gln Tyr Gly Thr Trp Tyr Lys Pro Glu Asn Ala Thr

405 410 415

Phe Val Asn Gly Asn Gln Pro Ile Val Thr Arg Ile Gly Ser Pro Phe

420 425 430

Leu Asn Ala Pro Val Gly Gly Asn Leu Pro Ala Gly Ala Thr Ile Val

435 440 445

Tyr Asp Glu Val Cys Ile Gln Ala Gly His Ile Trp Ile Gly Tyr Asn

450 455 460

Ala Tyr Asn Gly Asn Arg Val Tyr Cys Pro Val Arg Thr Cys Gln Gly

465 470 475 480

Val Pro Pro Asn Gln Ile Pro Gly Val Ala Trp Gly Val Phe Lys

485 490 495

<210> 24

<211> 281

<212> PROTEIN

<213>A118

<400> 24

Met Thr Ser Tyr Tyr Tyr Ser Arg Ser Leu Ala Asn Val Asn Lys Leu

1 5 10 15

Ala Asp Asn Thr Lys Ala Ala Ala Arg Lys Leu Leu Asp Trp Ser Glu

20 25 30

Ser Asn Gly Ile Glu Val Leu Ile Tyr Glu Thr Ile Arg Thr Lys Glu

35 40 45

Gln Gln Ala Ala Asn Val Asn Ser Gly Ala Ser Gln Thr Met Arg Ser

50 55 60

Tyr His Leu Val Gly Gln Ala Leu Asp Phe Val Met Ala Lys Gly Lys

65 70 75 80

Thr Val Asp Trp Gly Ala Tyr Arg Ser Asp Lys Gly Lys Lys Phe Val

85 90 95

Ala Lys Ala Lys Ser Leu Gly Phe Glu Trp Gly Gly Asp Trp Ser Gly

100 105 110

Phe Val Asp Asn Pro His Leu Gln Phe Asn Tyr Lys Gly Tyr Gly Thr

115 120 125

Asp Thr Phe Gly Lys Gly Ala Ser Thr Ser Asn Ser Ser Lys Pro Ser

130 135 140

Ala Asp Thr Asn Thr Asn Ser Leu Gly Leu Val Asp Tyr Met Asn Leu

145 150 155 160

Asn Lys Leu Asp Ser Ser Phe Ala Asn Arg Lys Lys Leu Ala Thr Ser

165 170 175

Tyr Gly Ile Lys Asn Tyr Ser Gly Thr Ala Thr Gln Asn Thr Thr Leu

180 185 190

Leu Ala Lys Leu Lys Ala Gly Lys Pro His Thr Pro Ala Ser Lys Asn

195 200 205

Thr Tyr Tyr Thr Glu Asn Pro Arg Lys Val Lys Thr Leu Val Gln Cys

210 215 220

Asp Leu Tyr Lys Ser Val Asp Phe Thr Thr Lys Asn Gln Thr Gly Gly

225 230 235 240

Thr Phe Pro Pro Gly Thr Val Phe Thr Ile Ser Gly Met Gly Lys Thr

245 250 255

Lys Gly Gly Thr Pro Arg Leu Lys Thr Lys Ser Gly Tyr Tyr Leu Thr

260 265 270

Ala Asn Thr Lys Phe Val Lys Lys Ile

275 280

<210> 25

<211> 341

<212> PROTEIN

<213>A511

<400> 25

Met Val Lys Tyr Thr Val Glu Asn Lys Ile Ile Ala Gly Leu Pro Lys

1 5 10 15

Gly Lys Leu Lys Gly Ala Asn Phe Val Ile Ala His Glu Thr Ala Asn

20 25 30

Ser Lys Ser Thr Ile Asp Asn Glu Val Ser Tyr Met Thr Arg Asn Trp

35 40 45

Lys Asn Ala Phe Val Thr His Phe Val Gly Gly Gly Gly Arg Val Val

50 55 60

Gln Val Ala Asn Val Asn Tyr Val Ser Trp Gly Ala Gly Gln Tyr Ala

65 70 75 80

Asn Ser Tyr Ser Tyr Ala Gln Val Glu Leu Cys Arg Thr Ser Asn Ala

85 90 95

Thr Thr Phe Lys Lys Asp Tyr Glu Val Tyr Cys Gln Leu Leu Val Asp

100 105 110

Leu Ala Lys Lys Ala Gly Ile Pro Ile Thr Leu Asp Ser Gly Ser Lys

115 120 125

Thr Ser Asp Lys Gly Ile Lys Ser His Lys Trp Val Ala Asp Lys Leu

130 135 140

Gly Gly Thr Thr His Gln Asp Pro Tyr Ala Tyr Leu Ser Ser Trp Gly

145 150 155 160

Ile Ser Lys Ala Gln Phe Ala Ser Asp Leu Ala Lys Val Ser Gly Gly

165 170 175

Gly Asn Thr Gly Thr Ala Pro Ala Lys Pro Ser Thr Pro Ala Pro Lys

180 185 190

Pro Ser Thr Pro Ser Thr Asn Leu Asp Lys Leu Gly Leu Val Asp Tyr

195 200 205

Met Asn Ala Lys Lys Met Asp Ser Ser Tyr Ser Asn Arg Asp Lys Leu

210 215 220

Ala Lys Gln Tyr Gly Ile Ala Asn Tyr Ser Gly Thr Ala Ser Gln Asn

225 230 235 240

Thr Thr Leu Leu Ser Lys Ile Lys Gly Gly Ala Pro Lys Pro Ser Thr

245 250 255

Pro Ala Pro Lys Pro Ser Thr Ser Thr Ala Lys Lys Ile Tyr Phe Pro

260 265 270

Pro Asn Lys Gly Asn Trp Ser Val Tyr Pro Thr Asn Lys Ala Pro Val

275 280 285

Lys Ala Asn Ala Ile Gly Ala Ile Asn Pro Thr Lys Phe Gly Gly Leu

290 295 300

Thr Tyr Thr Ile Gln Lys Asp Arg Gly Asn Gly Val Tyr Glu Ile Gln

305 310 315 320

Thr Asp Gln Phe Gly Arg Val Gln Val Tyr Gly Ala Pro Ser Thr Gly

325 330 335

Ala Val Ile Lys Lys

340

<210> 26

<211> 289

<212> PROTEIN

<213>A500

<400> 26

Met Ala Leu Thr Glu Ala Trp Leu Ile Glu Lys Ala Asn Arg Lys Leu

1 5 10 15

Asn Ala Gly Gly Met Tyr Lys Ile Thr Ser Asp Lys Thr Arg Asn Val

20 25 30

Ile Lys Lys Met Ala Lys Glu Gly Ile Tyr Leu Cys Val Ala Gln Gly

35 40 45

Tyr Arg Ser Thr Ala Glu Gln Asn Ala Leu Tyr Ala Gln Gly Arg Thr

50 55 60

Lys Pro Gly Ala Ile Val Thr Asn Ala Lys Gly Gly Gln Ser Asn His

65 70 75 80

Asn Tyr Gly Val Ala Val Asp Leu Cys Leu Tyr Thr Asn Asp Gly Lys

85 90 95

Asp Val Ile Trp Glu Ser Thr Thr Ser Arg Trp Lys Lys Val Val Ala

100 105 110

Ala Met Lys Ala Glu Gly Phe Lys Trp Gly Gly Asp Trp Lys Ser Phe

115 120 125

Lys Asp Tyr Pro His Phe Glu Leu Cys Asp Ala Val Ser Gly Glu Lys

130 135 140

Ile Pro Ala Ala Thr Gln Asn Thr Asn Thr Asn Ser Asn Arg Tyr Glu

145 150 155 160

Gly Lys Val Ile Asp Ser Ala Pro Leu Leu Pro Lys Met Asp Phe Lys

165 170 175

Ser Ser Pro Phe Arg Met Tyr Lys Val Gly Thr Glu Phe Leu Val Tyr

180 185 190

Asp His Asn Gln Tyr Trp Tyr Lys Thr Tyr Ile Asp Asp Lys Leu Tyr

195 200 205

Tyr Met Tyr Lys Ser Phe Cys Asp Val Val Ala Lys Lys Asp Ala Lys

210 215 220

Gly Arg Ile Lys Val Arg Ile Lys Ser Ala Lys Asp Leu Arg Ile Pro

225 230 235 240

Val Trp Asn Asn Ile Lys Leu Asn Ser Gly Lys Ile Lys Trp Tyr Ala

245 250 255

Pro Asn Val Lys Leu Ala Trp Tyr Asn Tyr Arg Arg Gly Tyr Leu Glu

260 265 270

Leu Trp Tyr Pro Asn Asp Gly Trp Tyr Tyr Thr Ala Glu Tyr Phe Leu

275 280 285

Lys

<210> 27

<211> 239

<212> PROTEIN

<213> Prophage LambdaSa1

<400> 27

Met Val Ile Asn Ile Glu Gln Ala Ile Ala Trp Met Ala Ser Arg Lys

1 5 10 15

Gly Lys Val Thr Tyr Ser Met Asp Tyr Arg Asn Gly Pro Ser Ser Tyr

20 25 30

Asp Cys Ser Ser Ser Val Tyr Phe Ala Leu Arg Ser Ala Gly Ala Ser

35 40 45

Asp Asn Gly Trp Ala Val Asn Thr Glu Tyr Glu His Asp Trp Leu Ile

50 55 60

Lys Asn Gly Tyr Val Leu Ile Ala Glu Asn Thr Asn Trp Asn Ala Gln

65 70 75 80

Arg Gly Asp Ile Phe Ile Trp Gly Lys Arg Gly Ala Ser Ala Gly Ala

85 90 95

Phe Gly His Thr Gly Met Phe Val Asp Pro Asp Asn Ile Ile His Cys

100 105 110

Asn Tyr Gly Tyr Asn Ser Ile Thr Val Asn Asn His Asp Glu Ile Trp

115 120 125

Gly Tyr Asn Gly Gln Pro Tyr Val Tyr Ala Tyr Arg Tyr Ala Arg Lys

130 135 140

Gln Ser Asn Ala Lys Val Asp Asn Gln Ser Val Val Ser Lys Phe Glu

145 150 155 160

Lys Glu Leu Asp Val Asn Thr Pro Leu Ser Asn Ser Asn Met Pro Tyr

165 170 175

Tyr Glu Ala Thr Ile Ser Glu Asp Tyr Tyr Val Glu Ser Lys Pro Asp

180 185 190

Val Asn Ser Thr Asp Lys Glu Leu Leu Val Ala Gly Thr Arg Val Arg

195 200 205

Val Tyr Glu Lys Val Lys Gly Trp Ala Arg Ile Gly Ala Pro Gln Ser

210 215 220

Asn Gln Trp Val Glu Asp Ala Tyr Leu Ile Asp Ala Thr Asp Met

225 230 235

<210> 28

<211> 468

<212> PROTEIN

<213> Prophage LambdaSa2

<400> 28

Met Glu Ile Asn Thr Glu Ile Ala Ile Ala Trp Met Ser Ala Arg Gln

1 5 10 15

Gly Lys Val Ser Tyr Ser Met Asp Tyr Arg Asp Gly Pro Asn Ser Tyr

20 25 30

Asp Cys Ser Ser Ser Val Tyr Tyr Ala Leu Arg Ser Ala Gly Ala Ser

35 40 45

Ser Ala Gly Trp Ala Val Asn Thr Glu Tyr Met His Asp Trp Leu Ile

50 55 60

Lys Asn Gly Tyr Glu Leu Ile Ala Glu Asn Val Asp Trp Asn Ala Val

65 70 75 80

Arg Gly Asp Ile Ala Ile Trp Gly Met Arg Gly His Ser Ser Gly Ala

85 90 95

Gly Gly His Val Val Met Phe Ile Asp Pro Glu Asn Ile Ile His Cys

100 105 110

Asn Trp Ala Asn Asn Gly Ile Thr Val Asn Asn Tyr Asn Gln Thr Ala

115 120 125

Ala Ala Ser Gly Trp Met Tyr Cys Tyr Val Tyr Arg Leu Lys Ser Gly

130 135 140

Ala Ser Thr Gln Gly Lys Ser Leu Asp Thr Leu Val Lys Glu Thr Leu

145 150 155 160

Ala Gly Asn Tyr Gly Asn Gly Glu Ala Arg Lys Ala Val Leu Gly Asn

165 170 175

Gln Tyr Glu Ala Val Met Ser Val Ile Asn Gly Lys Thr Thr Thr Asn

180 185 190

Gln Lys Thr Val Asp Gln Leu Val Gln Glu Val Ile Ala Gly Lys His

195 200 205

Gly Asn Gly Glu Ala Arg Lys Lys Ser Leu Gly Ser Gln Tyr Asp Ala

210 215 220

Val Gln Lys Arg Val Thr Glu Leu Leu Lys Lys Gln Pro Ser Glu Pro

225 230 235 240

Phe Lys Ala Gln Glu Val Asn Lys Pro Thr Glu Thr Lys Thr Ser Gln

245 250 255

Thr Glu Leu Thr Gly Gln Ala Thr Ala Thr Lys Glu Glu Gly Asp Leu

260 265 270

Ser Phe Asn Gly Thr Ile Leu Lys Lys Ala Val Leu Asp Lys Ile Leu

275 280 285

Gly Asn Cys Lys Lys His Asp Ile Leu Pro Ser Tyr Ala Leu Thr Ile

290 295 300

Leu His Tyr Glu Gly Leu Trp Gly Thr Ser Ala Val Gly Lys Ala Asp

305 310 315 320

Asn Asn Trp Gly Gly Met Thr Trp Thr Gly Gln Gly Asn Arg Pro Ser

325 330 335

Gly Val Thr Val Thr Gln Gly Ser Ala Arg Pro Ser Asn Glu Gly Gly

340 345 350

His Tyr Met His Tyr Ala Ser Val Asp Asp Phe Leu Thr Asp Trp Phe

355 360 365

Tyr Leu Leu Arg Ala Gly Gly Ser Tyr Lys Val Ser Gly Ala Lys Thr

370 375 380

Phe Ser Glu Ala Ile Lys Gly Met Phe Lys Val Gly Gly Ala Val Tyr

385 390 395 400

Asp Tyr Ala Ala Ser Gly Phe Asp Ser Tyr Ile Val Gly Ala Ser Ser

405 410 415

Arg Leu Lys Ala Ile Glu Ala Glu Asn Gly Ser Leu Asp Lys Phe Asp

420 425 430

Lys Ala Thr Asp Ile Gly Asp Gly Ser Lys Asp Lys Ile Asp Ile Thr

435 440 445

Ile Glu Gly Ile Glu Val Thr Ile Asn Gly Ile Thr Tyr Glu Leu Thr

450 455 460

Lys Lys Pro Val

465

<210> 29

<211> 236

<212> PROTEIN

<213> Prophage (ATCC700407)

<400> 29

Met Thr Asp Ser Ile Gln Glu Met Arg Lys Leu Gln Ser Ile Pro Val

1 5 10 15

Arg Tyr Asp Met Gly Asp Arg Tyr Gly Asn Asp Ala Asp Arg Asp Gly

20 25 30

Arg Ile Glu Met Asp Cys Ser Ser Ala Val Ser Lys Ala Leu Gly Ile

35 40 45

Ser Met Thr Asn Asn Thr Glu Thr Leu Gln Gln Ala Leu Pro Ala Ile

50 55 60

Gly Tyr Gly Lys Ile His Asp Ala Val Asp Gly Thr Phe Asp Met Gln

65 70 75 80

Ala Tyr Asp Val Ile Ile Trp Ala Pro Arg Asp Gly Ser Ser Ser Leu

85 90 95

Gly Ala Phe Gly His Val Leu Ile Ala Thr Ser Pro Thr Thr Ala Ile

100 105 110

His Cys Asn Tyr Gly Ser Asp Gly Ile Thr Glu Asn Asp Tyr Asn Tyr

115 120 125

Ile Trp Glu Ile Asn Gly Arg Pro Arg Glu Ile Val Phe Arg Lys Gly

130 135 140

Val Thr Gln Thr Gln Ala Thr Val Thr Ser Gln Phe Glu Arg Glu Leu

145 150 155 160

Asp Val Asn Ala Arg Leu Thr Val Ser Asp Lys Pro Tyr Tyr Glu Ala

165 170 175

Thr Leu Ser Glu Asp Tyr Tyr Val Glu Ala Gly Pro Arg Ile Asp Ser

180 185 190

Gln Asp Lys Glu Leu Ile Lys Ala Gly Thr Arg Val Arg Val Tyr Glu

195 200 205

Lys Leu Asn Gly Trp Ser Arg Ile Asn His Pro Glu Ser Ala Gln Trp

210 215 220

Val Glu Asp Ser Tyr Leu Val Asp Ala Thr Glu Met

225 230 235

<210> 30

<211> 481

<212> PROTEIN

<213> Phi11 and Phi12

<400> 30

Met Gln Ala Lys Leu Thr Lys Asn Glu Phe Ile Glu Trp Leu Lys Thr

1 5 10 15

Ser Glu Gly Lys Gln Phe Asn Val Asp Leu Trp Tyr Gly Phe Gln Cys

20 25 30

Phe Asp Tyr Ala Asn Ala Gly Trp Lys Val Leu Phe Gly Leu Leu Leu

35 40 45

Lys Gly Leu Gly Ala Lys Asp Ile Pro Phe Ala Asn Asn Phe Asp Gly

50 55 60

Leu Ala Thr Val Tyr Gln Asn Thr Pro Asp Phe Leu Ala Gln Pro Gly

65 70 75 80

Asp Met Val Val Phe Gly Ser Asn Tyr Gly Ala Gly Tyr Gly His Val

85 90 95

Ala Trp Val Ile Glu Ala Thr Leu Asp Tyr Ile Ile Val Tyr Glu Gln

100 105 110

Asn Trp Leu Gly Gly Gly Trp Thr Asp Gly Ile Glu Gln Pro Gly Trp

115 120 125

Gly Trp Glu Lys Val Thr Arg Arg Gln His Ala Tyr Asp Phe Pro Met

130 135 140

Trp Phe Ile Arg Pro Asn Phe Lys Ser Glu Thr Ala Pro Arg Ser Val

145 150 155 160

Gln Ser Pro Thr Gln Ala Pro Lys Lys Glu Thr Ala Lys Pro Gln Pro

165 170 175

Lys Ala Val Glu Leu Lys Ile Ile Lys Asp Val Val Lys Gly Tyr Asp

180 185 190

Leu Pro Lys Arg Gly Ser Asn Pro Lys Gly Ile Val Ile His Asn Asp

195 200 205

Ala Gly Ser Lys Gly Ala Thr Ala Glu Ala Tyr Arg Asn Gly Leu Val

210 215 220

Asn Ala Pro Leu Ser Arg Leu Glu Ala Gly Ile Ala His Ser Tyr Val

225 230 235 240

Ser Gly Asn Thr Val Trp Gln Ala Leu Asp Glu Ser Gln Val Gly Trp

245 250 255

His Thr Ala Asn Gln Ile Gly Asn Lys Tyr Tyr Tyr Gly Ile Glu Val

260 265 270

Cys Gln Ser Met Gly Ala Asp Asn Ala Thr Phe Leu Lys Asn Glu Gln

275 280 285

Ala Thr Phe Gln Glu Cys Ala Arg Leu Leu Lys Lys Trp Gly Leu Pro

290 295 300

Ala Asn Arg Asn Thr Ile Arg Leu His Asn Glu Phe Thr Ser Thr Ser

305 310 315 320

Cys Pro His Arg Ser Ser Val Leu His Thr Gly Phe Asp Pro Val Thr

325 330 335

Arg Gly Leu Leu Pro Glu Asp Lys Arg Leu Gln Leu Lys Asp Tyr Phe

340 345 350

Ile Lys Gln Ile Arg Ala Tyr Met Asp Gly Lys Ile Pro Val Ala Thr

355 360 365

Val Ser Asn Glu Ser Ser Ala Ser Ser Asn Thr Val Lys Pro Val Ala

370 375 380

Ser Ala Trp Lys Arg Asn Lys Tyr Gly Thr Tyr Tyr Met Glu Glu Ser

385 390 395 400

Ala Arg Phe Thr Asn Gly Asn Gln Pro Ile Thr Val Arg Lys Val Gly

405 410 415

Pro Phe Leu Ser Cys Pro Val Gly Tyr Gln Phe Gln Pro Gly Gly Tyr

420 425 430

Cys Asp Tyr Thr Glu Val Met Leu Gln Asp Gly His Val Trp Val Gly

435 440 445

Tyr Thr Trp Glu Gly Gln Arg Tyr Tyr Leu Pro Ile Arg Thr Trp Asn

450 455 460

Gly Ser Ala Pro Pro Asn Gln Ile Leu Gly Asp Leu Trp Gly Glu Ile

465 470 475 480

Ser

<210> 31

<211> 481

<212> PROTEIN

<213> (Phi)H5

<400> 31

Met Gln Ala Lys Leu Thr Lys Lys Glu Phe Ile Glu Trp Leu Lys Thr

1 5 10 15

Ser Glu Gly Lys Gln Tyr Asn Ala Asp Gly Trp Tyr Gly Phe Gln Cys

20 25 30

Phe Asp Tyr Ala Asn Ala Gly Trp Lys Ala Leu Phe Gly Leu Leu Leu

35 40 45

Lys Gly Val Gly Ala Lys Asp Ile Pro Phe Ala Asn Asn Phe Asp Gly

50 55 60

Leu Ala Thr Val Tyr Gln Asn Thr Pro Asp Phe Leu Ala Gln Pro Gly

65 70 75 80

Asp Met Val Val Phe Gly Ser Asn Tyr Gly Ala Gly Tyr Gly His Val

85 90 95

Ala Trp Val Ile Glu Ala Thr Leu Asp Tyr Ile Ile Val Tyr Glu Gln

100 105 110

Asn Trp Leu Gly Gly Gly Trp Thr Asp Gly Val Gln Gln Pro Gly Ser

115 120 125

Gly Trp Glu Lys Val Thr Arg Arg Gln His Ala Tyr Asp Phe Pro Met

130 135 140

Trp Phe Ile Arg Pro Asn Phe Lys Ser Glu Thr Ala Pro Arg Ser Val

145 150 155 160

Gln Ser Pro Thr Gln Ala Ser Lys Lys Glu Thr Ala Lys Pro Gln Pro

165 170 175

Lys Ala Val Glu Leu Lys Ile Ile Lys Asp Val Val Lys Gly Tyr Asp

180 185 190

Leu Pro Lys Arg Gly Ser Asn Pro Asn Phe Ile Val Ile His Asn Asp

195 200 205

Ala Gly Ser Lys Gly Ala Thr Ala Glu Ala Tyr Arg Asn Gly Leu Val

210 215 220

Asn Ala Pro Leu Ser Arg Leu Glu Ala Gly Ile Ala His Ser Tyr Val

225 230 235 240

Ser Gly Asn Thr Val Trp Gln Ala Leu Asp Glu Ser Gln Val Gly Trp

245 250 255

His Thr Ala Asn Gln Ile Gly Asn Lys Tyr Gly Tyr Gly Ile Glu Val

260 265 270

Cys Gln Ser Met Gly Ala Asp Asn Ala Thr Phe Leu Lys Asn Glu Gln

275 280 285

Ala Thr Phe Gln Glu Cys Ala Arg Leu Leu Lys Lys Trp Gly Leu Pro

290 295 300

Ala Asn Arg Asn Thr Ile Arg Leu His Asn Glu Phe Thr Ser Thr Ser

305 310 315 320

Cys Pro His Arg Ser Ser Val Leu His Thr Gly Phe Asp Pro Val Thr

325 330 335

Arg Gly Leu Leu Pro Glu Asp Lys Arg Leu Gln Leu Lys Asp Tyr Phe

340 345 350

Ile Lys Gln Ile Arg Ala Tyr Met Asp Gly Lys Ile Pro Val Ala Thr

355 360 365

Val Ser Asn Asp Ser Ser Ala Ser Ser Asn Thr Val Lys Pro Val Ala

370 375 380

Ser Ala Trp Lys Arg Asn Lys Tyr Gly Thr Tyr Tyr Met Glu Glu Ser

385 390 395 400

Ala Arg Phe Thr Asn Gly Asn Gln Pro Ile Thr Val Arg Lys Val Gly

405 410 415

Pro Phe Leu Ser Cys Pro Val Gly Tyr Gln Phe Gln Pro Gly Gly Tyr

420 425 430

Cys Asp Tyr Thr Glu Val Met Leu Gln Asp Gly His Val Trp Val Gly

435 440 445

Tyr Thr Trp Glu Gly Gln Arg Tyr Tyr Leu Pro Ile Arg Thr Trp Asn

450 455 460

Gly Ser Ala Pro Pro Asn Gln Ile Leu Gly Asp Leu Trp Gly Glu Ile

465 470 475 480

Ser

<210> 32

<211> 477

<212> PROTEIN

<213> (Phi)WMY

<400> 32

Met Lys Thr Lys Ala Gln Ala Lys Ser Trp Ile Asn Ser Lys Ile Gly

1 5 10 15

Lys Gly Ile Asp Trp Asp Gly Met Tyr Gly Tyr Gln Cys Met Asp Glu

20 25 30

Ala Val Asp Tyr Ile His His Val Thr Asp Gly Lys Val Thr Met Trp

35 40 45

Gly Asn Ala Ile Asp Ala Pro Lys Asn Asn Phe Gln Gly Leu Cys Thr

50 55 60

Val Tyr Thr Asn Thr Pro Glu Phe Arg Pro Ala Tyr Gly Asp Val Ile

65 70 75 80

Val Trp Ser Tyr Gly Thr Phe Ala Thr Tyr Gly His Ile Ala Ile Val

85 90 95

Val Asn Pro Asp Pro Tyr Gly Asp Leu Gln Tyr Ile Thr Val Leu Glu

100 105 110

Gln Asn Trp Asn Gly Asn Gly Ile Tyr Lys Thr Glu Phe Ala Thr Ile

115 120 125

Arg Thr His Asp Tyr Thr Gly Val Ser His Phe Ile Arg Pro Lys Phe

130 135 140

Ala Asp Glu Val Lys Glu Thr Ala Lys Thr Val Asn Lys Leu Ser Val

145 150 155 160

Gln Lys Lys Ile Val Thr Pro Lys Asn Ser Val Glu Arg Ile Lys Asn

165 170 175

Tyr Val Lys Thr Ser Gly Tyr Ile Asn Gly Glu His Tyr Glu Leu Tyr

180 185 190

Asn Arg Gly His Lys Pro Lys Gly Val Val Ile His Asn Thr Ala Gly

195 200 205

Thr Ala Ser Ala Thr Gln Glu Gly Gln Arg Leu Thr Asn Met Thr Phe

210 215 220

Gln Gln Leu Ala Asn Gly Val Ala His Val Tyr Ile Asp Lys Asn Thr

225 230 235 240

Ile Tyr Glu Thr Leu Pro Glu Asp Arg Ile Ala Trp His Val Ala Gln

245 250 255

Gln Tyr Gly Asn Thr Glu Phe Tyr Gly Ile Glu Val Cys Gly Ser Arg

260 265 270

Asn Thr Asp Lys Glu Gln Phe Leu Ala Asn Glu Gln Val Ala Phe Gln

275 280 285

Glu Ala Ala Arg Arg Leu Lys Ser Trp Gly Met Arg Ala Asn Arg Asn

290 295 300

Thr Val Arg Leu His His Thr Phe Ser Ser Thr Glu Cys Pro Asp Met

305 310 315 320

Ser Met Leu Leu His Thr Gly Tyr Ser Met Lys Asn Gly Lys Pro Thr

325 330 335

Gln Asp Ile Thr Asn Lys Cys Ala Asp Tyr Phe Met Lys Gln Ile Asn

340 345 350

Ala Tyr Ile Asp Gly Lys Gln Pro Thr Ser Thr Val Val Gly Ser Ser

355 360 365

Ser Ser Asn Lys Leu Lys Ala Lys Asn Lys Asp Lys Ser Thr Gly Trp

370 375 380

Asn Thr Asn Glu Tyr Gly Thr Leu Trp Lys Lys Glu His Ala Thr Phe

385 390 395 400

Thr Cys Gly Val Arg Gln Gly Ile Val Thr Arg Thr Thr Gly Pro Phe

405 410 415

Thr Ser Cys Pro Gln Ala Gly Val Leu Tyr Tyr Gly Gln Ser Val Asn

420 425 430

Tyr Asp Thr Val Cys Lys Gln Asp Gly Tyr Val Trp Ile Ser Trp Thr

435 440 445

Thr Ser Asp Gly Tyr Asp Val Trp Met Pro Ile Arg Thr Trp Asp Arg

450 455 460

Ser Thr Asp Lys Val Ser Glu Ile Trp Gly Thr Ile Ser

465 470 475

<210> 33

<211> 443

<212> PROTEIN

<213> (Phi)NCTC 11261

<400> 33

Met Ala Thr Tyr Gln Glu Tyr Lys Ser Arg Ser Asn Gly Asn Ala Tyr

1 5 10 15

Asp Ile Asp Gly Ser Phe Gly Ala Gln Cys Trp Asp Gly Tyr Ala Asp

20 25 30

Tyr Cys Lys Tyr Leu Gly Leu Pro Tyr Ala Asn Cys Thr Asn Thr Gly

35 40 45

Tyr Ala Arg Asp Ile Trp Glu Gln Arg His Glu Asn Gly Ile Leu Asn

50 55 60

Tyr Phe Asp Glu Val Glu Val Met Gln Ala Gly Asp Val Ala Ile Phe

65 70 75 80

Met Val Val Asp Gly Val Thr Pro Tyr Ser His Val Ala Ile Phe Asp

85 90 95

Ser Asp Ala Gly Gly Gly Tyr Gly Trp Phe Leu Gly Gln Asn Gln Gly

100 105 110

Gly Ala Asn Gly Ala Tyr Asn Ile Val Lys Ile Pro Tyr Ser Ala Thr

115 120 125

Tyr Pro Thr Ala Phe Arg Pro Lys Val Phe Lys Asn Ala Val Thr Val

130 135 140

Thr Gly Asn Ile Gly Leu Asn Lys Gly Asp Tyr Phe Ile Asp Val Ser

145 150 155 160

Ala Tyr Gln Gln Ala Asp Leu Thr Thr Thr Cys Gln Gln Ala Gly Thr

165 170 175

Thr Lys Thr Ile Ile Lys Val Ser Glu Ser Ile Ala Trp Leu Ser Asp

180 185 190

Arg His Gln Gln Gln Ala Asn Thr Ser Asp Pro Ile Gly Tyr Tyr His

195 200 205

Phe Gly Arg Phe Gly Gly Asp Ser Ala Leu Ala Gln Arg Glu Ala Asp

210 215 220

Leu Phe Leu Ser Asn Leu Pro Ser Lys Lys Val Ser Tyr Leu Val Ile

225 230 235 240

Asp Tyr Glu Asp Ser Ala Ser Ala Asp Lys Gln Ala Asn Thr Asn Ala

245 250 255

Val Ile Ala Phe Met Asp Lys Ile Ala Ser Ala Gly Tyr Lys Pro Ile

260 265 270

Tyr Tyr Ser Tyr Lys Pro Phe Thr Leu Asn Asn Ile Asp Tyr Gln Lys

275 280 285

Ile Ile Ala Lys Tyr Pro Asn Ser Ile Trp Ile Ala Gly Tyr Pro Asp

290 295 300

Tyr Glu Val Arg Thr Glu Pro Leu Trp Glu Phe Phe Pro Ser Met Asp

305 310 315 320

Gly Val Arg Trp Trp Gln Phe Thr Ser Val Gly Val Ala Gly Gly Leu

325 330 335

Asp Lys Asn Ile Val Leu Leu Ala Asp Asp Ser Ser Lys Met Asp Ile

340 345 350

Pro Lys Val Asp Lys Pro Gln Glu Leu Thr Phe Tyr Gln Lys Leu Ala

355 360 365

Thr Asn Thr Lys Leu Asp Asn Ser Asn Val Pro Tyr Tyr Glu Ala Thr

370 375 380

Leu Ser Thr Asp Tyr Tyr Val Glu Ser Lys Pro Asn Ala Ser Ser Ala

385 390 395 400

Asp Lys Glu Phe Ile Lys Ala Gly Thr Arg Val Arg Val Tyr Glu Lys

405 410 415

Val Asn Gly Trp Ser Arg Ile Asn His Pro Glu Ser Ala Gln Trp Val

420 425 430

Glu Asp Ser Tyr Leu Val Asn Ala Thr Asp Met

435 440

<210> 34

<211> 334

<212> PROTEIN

<213> (Phi)FWLLm3

<400> 34

Met Val Lys Tyr Thr Val Glu Asn Lys Ile Ile Ala Gly Leu Pro Lys

1 5 10 15

Gly Lys Leu Lys Gly Ala Asn Phe Val Ile Ala His Glu Thr Ala Asn

20 25 30

Ser Lys Ser Thr Ile Asp Asn Glu Val Ser Tyr Met Thr Arg Asn Trp

35 40 45

Gln Asn Ala Phe Val Thr His Phe Val Gly Gly Gly Gly Arg Val Val

50 55 60

Gln Val Ala Asn Val Asn Tyr Val Ser Trp Gly Ala Gly Gln Tyr Ala

65 70 75 80

Asn Ser Tyr Ser Tyr Ala Gln Val Glu Leu Cys Arg Thr Ser Asn Ala

85 90 95

Thr Thr Phe Lys Lys Asp Tyr Glu Val Tyr Cys Gln Leu Leu Val Asp

100 105 110

Leu Ala Lys Lys Ala Gly Ile Pro Ile Thr Leu Asp Ser Gly Ser Lys

115 120 125

Thr Ser Asp Lys Gly Ile Lys Ser His Lys Trp Val Ala Asp Lys Leu

130 135 140

Gly Gly Thr Thr His Gln Asp Pro Tyr Ala Tyr Leu Ser Ser Trp Gly

145 150 155 160

Ile Ser Lys Ala Gln Phe Ala Ser Asp Leu Ala Lys Val Ser Gly Gly

165 170 175

Gly Asn Thr Gly Thr Ala Pro Ala Lys Pro Ser Thr Pro Ser Thr Asn

180 185 190

Leu Asp Lys Leu Gly Leu Val Asp Tyr Met Asn Ala Lys Lys Met Asp

195 200 205

Ser Ser Tyr Ser Asn Arg Ala Lys Leu Ala Lys Gln Tyr Gly Ile Ala

210 215 220

Asn Tyr Ser Gly Thr Ala Ser Gln Asn Thr Thr Leu Leu Ser Lys Ile

225 230 235 240

Lys Gly Gly Ala Pro Lys Pro Ser Thr Pro Ala Pro Lys Pro Ser Thr

245 250 255

Ser Thr Ala Lys Lys Ile Tyr Phe Pro Pro Asn Lys Gly Asn Trp Ser

260 265 270

Val Tyr Pro Thr Asn Lys Ala Pro Val Lys Ala Asn Ala Ile Gly Ala

275 280 285

Ile Asn Pro Thr Lys Phe Gly Gly Leu Thr Tyr Thr Ile Gln Lys Asp

290 295 300

Arg Gly Asn Gly Val Tyr Glu Ile Gln Thr Asp Gln Phe Gly Arg Val

305 310 315 320

Gln Val Tyr Gly Ala Pro Ser Thr Gly Ala Val Ile Lys Lys

325 330

<210> 35

<211> 1278

<212> PROTEIN

<213> (Phi)BPS13

<400> 35

Met Ala Lys Arg Glu Lys Tyr Ile Phe Asp Val Glu Ala Glu Val Gly

1 5 10 15

Lys Ala Ala Lys Ser Ile Lys Ser Leu Glu Ala Glu Leu Ser Lys Leu

20 25 30

Gln Lys Leu Asn Lys Glu Ile Asp Ala Thr Gly Gly Asp Arg Thr Glu

35 40 45

Lys Glu Met Leu Ala Thr Leu Lys Ala Ala Lys Glu Val Asn Ala Glu

50 55 60

Tyr Gln Lys Met Gln Arg Ile Leu Lys Asp Leu Ser Lys Tyr Ser Gly

65 70 75 80

Lys Val Ser Arg Lys Glu Phe Asn Asp Ser Lys Val Ile Asn Asn Ala

85 90 95

Lys Thr Ser Val Gln Gly Gly Lys Val Thr Asp Ser Phe Gly Gln Met

100 105 110

Leu Lys Asn Met Glu Arg Gln Ile Asn Ser Val Asn Lys Gln Phe Asp

115 120 125

Asn His Arg Lys Ala Met Val Asp Arg Gly Gln Gln Tyr Thr Pro His

130 135 140

Leu Lys Thr Asn Arg Lys Asp Ser Gln Gly Asn Ser Asn Pro Ser Met

145 150 155 160

Met Gly Arg Asn Lys Ser Thr Thr Gln Asp Met Glu Lys Ala Val Asp

165 170 175

Lys Phe Leu Asn Gly Gln Asn Glu Ala Thr Thr Gly Leu Asn Gln Ala

180 185 190

Leu Tyr Gln Leu Lys Glu Ile Ser Lys Leu Asn Arg Arg Ser Glu Ser

195 200 205

Leu Ser Arg Arg Ala Ser Ala Ser Gly Tyr Met Ser Phe Gln Gln Tyr

210 215 220

Ser Asn Phe Thr Gly Asp Arg Arg Thr Val Gln Gln Thr Tyr Gly Gly

225 230 235 240

Leu Lys Thr Ala Asn Arg Glu Arg Val Leu Glu Leu Ser Gly Gln Ala

245 250 255

Thr Gly Ile Ser Lys Glu Leu Asp Arg Leu Asn Ser Lys Lys Gly Leu

260 265 270

Thr Ala Arg Glu Gly Glu Glu Arg Lys Lys Leu Met Arg Gln Leu Glu

275 280 285

Gly Ile Asp Ala Glu Leu Thr Ala Arg Lys Lys Leu Asn Ser Ser Leu

290 295 300

Asp Glu Thr Thr Ser Asn Met Glu Lys Phe Asn Gln Ser Leu Val Asp

305 310 315 320

Ala Gln Val Ser Val Lys Pro Glu Arg Gly Thr Met Arg Gly Met Met

325 330 335

Tyr Glu Arg Ala Pro Ala Ile Ala Leu Ala Ile Gly Gly Ala Ile Thr

340 345 350

Ala Thr Ile Gly Lys Leu Tyr Ser Glu Gly Gly Asn His Ser Lys Ala

355 360 365

Met Arg Pro Asp Glu Met Tyr Val Gly Gln Gln Thr Gly Ala Val Gly

370 375 380

Ala Asn Trp Arg Pro Asn Arg Thr Ala Thr Met Arg Ser Gly Leu Gly

385 390 395 400

Asn His Leu Gly Phe Thr Gly Gln Glu Met Met Glu Phe Gln Ser Asn

405 410 415

Tyr Leu Ser Ala Asn Gly Tyr His Gly Ala Glu Asp Met Lys Ala Ala

420 425 430

Thr Thr Gly Gln Ala Thr Phe Ala Arg Ala Thr Gly Leu Gly Ser Asp

435 440 445

Glu Val Lys Asp Phe Phe Asn Thr Ala Tyr Arg Ser Gly Gly Ile Asp

450 455 460

Gly Asn Gln Thr Lys Gln Phe Gln Asn Ala Phe Leu Gly Ala Met Lys

465 470 475 480

Gln Ser Gly Ala Val Gly Arg Glu Lys Asp Gln Leu Lys Ala Leu Asn

485 490 495

Gly Ile Leu Ser Ser Met Ser Gln Asn Arg Thr Val Ser Asn Gln Asp

500 505 510

Met Met Arg Thr Val Gly Leu Gln Ser Ala Ile Ser Ser Ser Gly Val

515 520 525

Ala Ser Leu Gln Gly Thr Lys Gly Gly Ala Leu Met Glu Gln Leu Asp

530 535 540

Asn Gly Ile Arg Glu Gly Phe Asn Asp Pro Gln Met Arg Val Leu Phe

545 550 555 560

Gly Gln Gly Thr Lys Tyr Gln Gly Met Gly Gly Arg Ala Ala Leu Arg

565 570 575

Lys Gln Met Glu Lys Gly Ile Ser Asp Pro Asp Asn Leu Asn Thr Leu

580 585 590

Ile Asp Ala Ser Lys Ala Ser Ala Gly Gln Asp Pro Ala Glu Gln Ala

595 600 605

Glu Val Leu Ala Thr Leu Ala Ser Lys Met Gly Val Asn Met Ser Ser

610 615 620

Asp Gln Ala Arg Gly Leu Ile Asp Leu Gln Pro Ser Gly Lys Leu Thr

625 630 635 640

Lys Glu Asn Ile Asp Lys Val Met Lys Glu Gly Leu Lys Glu Gly Ser

645 650 655

Ile Glu Ser Ala Lys Arg Asp Lys Ala Tyr Ser Glu Ser Lys Ala Ser

660 665 670

Ile Asp Asn Ser Ser Glu Ala Ala Thr Ala Lys Gln Ala Thr Glu Leu

675 680 685

Asn Asp Met Gly Ser Lys Leu Arg Gln Ala Asn Ala Ala Leu Gly Gly

690 695 700

Leu Pro Ala Pro Leu Tyr Thr Ala Ile Ala Ala Val Val Ala Phe Thr

705 710 715 720

Ala Ala Val Ala Gly Ser Ala Leu Met Phe Lys Gly Ala Ser Trp Leu

725 730 735

Lys Gly Gly Met Ala Ser Lys Tyr Gly Gly Lys Gly Gly Lys Gly Gly

740 745 750

Lys Gly Gly Gly Thr Gly Gly Gly Gly Gly Ala Gly Gly Ala Ala Ala

755 760 765

Thr Gly Ala Gly Ala Ala Ala Gly Ala Gly Gly Val Gly Ala Ala Ala

770 775 780

Ala Gly Glu Val Gly Ala Gly Val Ala Ala Gly Gly Ala Ala Ala Gly

785 790 795 800

Ala Ala Ala Gly Gly Ser Lys Leu Ala Gly Val Gly Lys Gly Phe Met

805 810 815

Lys Gly Ala Gly Lys Leu Met Leu Pro Leu Gly Ile Leu Met Gly Ala

820 825 830

Ser Glu Ile Met Gln Ala Pro Glu Glu Ala Lys Gly Ser Ala Ile Gly

835 840 845

Ser Ala Val Gly Gly Ile Gly Gly Gly Ile Ala Gly Gly Ala Ala Thr

850 855 860

Gly Ala Ile Ala Gly Ser Phe Leu Gly Pro Ile Gly Thr Ala Val Gly

865 870 875 880

Gly Ile Ala Gly Gly Ile Ala Gly Gly Phe Ala Gly Ser Ser Leu Gly

885 890 895

Glu Thr Ile Gly Gly Trp Phe Asp Ser Gly Pro Lys Glu Asp Ala Ser

900 905 910

Ala Ala Asp Lys Ala Lys Ala Asp Ala Ser Ala Ala Ala Leu Ala Ala

915 920 925

Ala Ala Gly Thr Ser Gly Ala Val Gly Ser Ser Ala Leu Gln Ser Gln

930 935 940

Met Ala Gln Gly Ile Thr Gly Ala Pro Asn Met Ser Gln Val Gly Ser

945 950 955 960

Met Ala Ser Ala Leu Gly Ile Ser Ser Gly Ala Met Ala Ser Ala Leu

965 970 975

Gly Ile Ser Ser Gly Gln Glu Asn Gln Ile Gln Thr Met Thr Asp Lys

980 985 990

Glu Asn Thr Asn Thr Lys Lys Ala Asn Glu Ala Lys Lys Gly Asp Asn

995 1000 1005

Leu Ser Tyr Glu Arg Glu Asn Ile Ser Met Tyr Glu Arg Val Leu

1010 1015 1020

Thr Arg Ala Glu Gln Ile Leu Ala Gln Ala Arg Ala Gln Asn Gly

1025 1030 1035

Ile Met Gly Val Gly Gly Gly Gly Thr Ala Gly Ala Gly Gly Gly

1040 1045 1050

Ile Asn Gly Phe Thr Gly Gly Gly Lys Leu Gln Phe Leu Ala Ala

1055 1060 1065

Gly Gln Lys Trp Ser Ser Ser Asn Leu Gln Gln His Asp Leu Gly

1070 1075 1080

Phe Thr Asp Gln Asn Leu Thr Ala Glu Asp Leu Asp Lys Trp Ile

1085 1090 1095

Asp Ser Lys Ala Pro Gln Gly Ser Met Met Arg Gly Met Gly Ala

1100 1105 1110

Thr Phe Leu Lys Ala Gly Gln Glu Tyr Gly Leu Asp Pro Arg Tyr

1115 1120 1125

Leu Ile Ala His Ala Ala Glu Glu Ser Gly Trp Gly Thr Ser Lys

1130 1135 1140

Ile Ala Arg Asp Lys Gly Asn Phe Phe Gly Ile Gly Ala Phe Asp

1145 1150 1155

Asp Ser Pro Tyr Ser Ser Ala Tyr Glu Phe Lys Asp Gly Thr Gly

1160 1165 1170

Ser Ala Ala Glu Arg Gly Ile Met Gly Gly Ala Lys Trp Ile Ser

1175 1180 1185

Glu Lys Tyr Tyr Gly Lys Gly Asn Thr Thr Leu Asp Lys Met Lys

1190 1195 1200

Ala Ala Gly Tyr Ala Thr Asn Ala Ser Trp Ala Pro Asn Ile Ala

1205 1210 1215

Ser Ile Met Ala Gly Ala Pro Thr Gly Ser Gly Ser Gly Asn Val

1220 1225 1230

Thr Ala Thr Ile Asn Val Asn Val Lys Gly Asp Glu Lys Val Ser

1235 1240 1245

Asp Lys Leu Lys Asn Ser Ser Asp Met Lys Lys Ala Gly Lys Asp

1250 1255 1260

Ile Gly Ser Leu Leu Gly Phe Tyr Ser Arg Glu Met Thr Ile Ala

1265 1270 1275

<210> 36

<211> 495

<212> PROTEIN

<213> (Phi)GH15

<400> 36

Met Ala Lys Thr Gln Ala Glu Ile Asn Lys Arg Leu Asp Ala Tyr Ala

1 5 10 15

Lys Gly Thr Val Asp Ser Pro Tyr Arg Ile Lys Lys Ala Thr Ser Tyr

20 25 30

Asp Pro Ser Phe Gly Val Met Glu Ala Gly Ala Ile Asp Ala Asp Gly

35 40 45

Tyr Tyr His Ala Gln Cys Gln Asp Leu Ile Thr Asp Tyr Val Leu Trp

50 55 60

Leu Thr Asp Asn Lys Val Arg Thr Trp Gly Asn Ala Lys Asp Gln Ile

65 70 75 80

Lys Gln Ser Tyr Gly Thr Gly Phe Lys Ile His Glu Asn Lys Pro Ser

85 90 95

Thr Val Pro Lys Lys Gly Trp Ile Ala Val Phe Thr Ser Gly Ser Tyr

100 105 110

Gln Gln Trp Gly His Ile Gly Ile Val Tyr Asp Gly Gly Asn Thr Ser

115 120 125

Thr Phe Thr Ile Leu Glu Gln Asn Trp Asn Gly Tyr Ala Asn Lys Lys

130 135 140

Pro Thr Lys Arg Val Asp Asn Tyr Tyr Gly Leu Thr His Phe Ile Glu

145 150 155 160

Ile Pro Val Lys Ala Gly Thr Thr Val Lys Lys Glu Thr Ala Lys Lys

165 170 175

Ser Ala Ser Lys Thr Pro Ala Pro Lys Lys Lys Ala Thr Leu Lys Val

180 185 190

Ser Lys Asn His Ile Asn Tyr Thr Met Asp Lys Arg Gly Lys Lys Pro

195 200 205

Glu Gly Met Val Ile His Asn Asp Ala Gly Arg Ser Ser Gly Gln Gln

210 215 220

Tyr Glu Asn Ser Leu Ala Asn Ala Gly Tyr Ala Arg Tyr Ala Asn Gly

225 230 235 240

Ile Ala His Tyr Tyr Gly Ser Glu Gly Tyr Val Trp Glu Ala Ile Asp

245 250 255

Ala Lys Asn Gln Ile Ala Trp His Thr Gly Asp Gly Thr Gly Ala Asn

260 265 270

Ser Gly Asn Phe Arg Phe Ala Gly Ile Glu Val Cys Gln Ser Met Ser

275 280 285

Ala Ser Asp Ala Gln Phe Leu Lys Asn Glu Gln Ala Val Phe Gln Phe

290 295 300

Thr Ala Glu Lys Phe Lys Glu Trp Gly Leu Thr Pro Asn Arg Lys Thr

305 310 315 320

Val Arg Leu His Met Glu Phe Val Pro Thr Ala Cys Pro His Arg Ser

325 330 335

Met Val Leu His Thr Gly Phe Asn Pro Val Thr Gln Gly Arg Pro Ser

340 345 350

Gln Ala Ile Met Asn Lys Leu Lys Asp Tyr Phe Ile Lys Gln Ile Lys

355 360 365

Asn Tyr Met Asp Lys Gly Thr Ser Ser Ser Thr Val Val Lys Asp Gly

370 375 380

Lys Thr Ser Ser Ala Ser Thr Pro Ala Thr Arg Pro Val Thr Gly Ser

385 390 395 400

Trp Lys Lys Asn Gln Tyr Gly Thr Trp Tyr Lys Pro Glu Asn Ala Thr

405 410 415

Phe Val Asn Gly Asn Gln Pro Ile Val Thr Arg Ile Gly Ser Pro Phe

420 425 430

Leu Asn Ala Pro Val Gly Gly Asn Leu Pro Ala Gly Ala Thr Ile Val

435 440 445

Tyr Asp Glu Val Cys Ile Gln Ala Gly His Ile Trp Ile Gly Tyr Asn

450 455 460

Ala Tyr Asn Gly Asp Arg Val Tyr Cys Pro Val Arg Thr Cys Gln Gly

465 470 475 480

Val Pro Pro Asn His Ile Pro Gly Val Ala Trp Gly Val Phe Lys

485 490 495

<210> 37

<211> 264

<212> PROTEIN

<213> (Phi)8074-B1

<400> 37

Met Lys Ile Gly Ile Asp Met Gly His Thr Leu Ser Gly Ala Asp Tyr

1 5 10 15

Gly Val Val Gly Leu Arg Pro Glu Ser Val Leu Thr Arg Glu Val Gly

20 25 30

Thr Lys Val Ile Tyr Lys Leu Gln Lys Leu Gly His Val Val Val Asn

35 40 45

Cys Thr Val Asp Lys Ala Ser Ser Val Ser Glu Ser Leu Tyr Thr Arg

50 55 60

Tyr Tyr Arg Ala Asn Gln Ala Asn Val Asp Leu Phe Ile Ser Ile His

65 70 75 80

Phe Asn Ala Thr Pro Gly Gly Thr Gly Thr Glu Val Tyr Thr Tyr Ala

85 90 95

Gly Arg Gln Leu Gly Glu Ala Thr Arg Ile Arg Gln Glu Phe Lys Ser

100 105 110

Leu Gly Leu Arg Asp Arg Gly Thr Lys Asp Gly Ser Gly Leu Ala Val

115 120 125

Ile Arg Asn Thr Lys Ala Lys Ala Met Leu Val Glu Cys Cys Phe Cys

130 135 140

Asp Asn Pro Asn Asp Met Lys Leu Tyr Asn Ser Glu Ser Phe Ser Asn

145 150 155 160

Ala Ile Val Lys Gly Ile Thr Gly Lys Leu Pro Asn Gly Glu Ser Gly

165 170 175

Asn Asn Asn Gln Gly Gly Asn Lys Val Lys Ala Val Val Ile Tyr Asn

180 185 190

Glu Gly Ala Asp Arg Arg Gly Ala Glu Tyr Leu Ala Asp Tyr Leu Asn

195 200 205

Cys Pro Thr Ile Ser Asn Ser Arg Thr Phe Asp Tyr Ser Cys Val Glu

210 215 220

His Val Tyr Ala Val Gly Gly Lys Lys Glu Gln Tyr Thr Lys Tyr Leu

225 230 235 240

Lys Thr Leu Leu Ser Gly Ala Asn Arg Tyr Asp Thr Met Gln Gln Ile

245 250 255

Leu Asn Phe Ile Asn Gly Gly Lys

260

<210> 38

<211> 209

<212> PROTEIN

<213> (Phi)SPN1S

<400> 38

Met Asp Ile Asn Gln Phe Arg Arg Ala Ser Gly Ile Asn Glu Gln Leu

1 5 10 15

Ala Ala Arg Trp Phe Pro His Ile Thr Thr Ala Met Asn Glu Phe Gly

20 25 30

Ile Thr Lys Pro Asp Asp Gln Ala Met Phe Ile Ala Gln Val Gly His

35 40 45

Glu Ser Gly Gly Phe Thr Arg Leu Gln Glu Asn Phe Asn Tyr Ser Val

50 55 60

Asn Gly Leu Ser Gly Phe Ile Arg Ala Gly Arg Ile Thr Pro Asp Gln

65 70 75 80

Ala Asn Ala Leu Gly Arg Lys Thr Tyr Glu Lys Ser Leu Pro Leu Glu

85 90 95

Arg Gln Arg Ala Ile Ala Asn Leu Val Tyr Ser Lys Arg Met Gly Asn

100 105 110

Asn Gly Pro Gly Asp Gly Trp Asn Tyr Arg Gly Arg Gly Leu Ile Gln

115 120 125

Ile Thr Gly Leu Asn Asn Tyr Arg Asp Cys Gly Asn Gly Leu Lys Val

130 135 140

Asp Leu Val Ala Gln Pro Glu Leu Leu Ala Gln Asp Glu Tyr Ala Ala

145 150 155 160

Arg Ser Ala Ala Trp Phe Phe Ser Ser Lys Gly Cys Met Lys Tyr Thr

165 170 175

Gly Asp Leu Val Arg Val Thr Gln Ile Ile Asn Gly Gly Gln Asn Gly

180 185 190

Ile Asp Asp Arg Arg Thr Arg Tyr Ala Ala Ala Arg Lys Val Leu Ala

195 200 205

Leu

<210> 39

<211> 290

<212> PROTEIN

<213> (Phi)CN77

<400> 39

Met Gly Tyr Trp Gly Tyr Pro Asn Gly Gln Ile Pro Asn Asp Lys Met

1 5 10 15

Ala Leu Tyr Arg Gly Cys Leu Leu Arg Ala Asp Ala Ala Ala Gln Ala

20 25 30

Tyr Ala Leu Gln Asp Ala Tyr Thr Arg Ala Thr Gly Lys Pro Leu Val

35 40 45

Ile Leu Glu Gly Tyr Arg Asp Leu Thr Arg Gln Lys Tyr Leu Arg Asn

50 55 60

Leu Tyr Leu Ser Gly Arg Gly Asn Ile Ala Ala Val Pro Gly Leu Ser

65 70 75 80

Asn His Gly Trp Gly Leu Ala Cys Asp Phe Ala Ala Pro Leu Asn Ser

85 90 95

Ser Gly Ser Glu Glu His Arg Trp Met Arg Gln Asn Ala Pro Leu Phe

100 105 110

Gly Phe Asp Trp Ala Arg Gly Lys Ala Asp Asn Glu Pro Trp His Trp

115 120 125

Glu Tyr Gly Asn Val Pro Val Ser Arg Trp Ala Ser Leu Asp Val Thr

130 135 140

Pro Ile Asp Arg Asn Asp Met Ala Asp Ile Thr Glu Gly Gln Met Gln

145 150 155 160

Arg Ile Ala Val Ile Leu Leu Asp Thr Glu Ile Gln Thr Pro Leu Gly

165 170 175

Pro Arg Leu Val Lys His Ala Leu Gly Asp Ala Leu Leu Leu Gly Gln

180 185 190

Ala Asn Ala Asn Ser Ile Ala Glu Val Pro Asp Lys Thr Trp Asp Val

195 200 205

Leu Val Asp His Pro Leu Ala Lys Asn Glu Asp Gly Thr Pro Leu Lys

210 215 220

Val Arg Leu Gly Asp Val Ala Lys Tyr Glu Pro Leu Glu His Gln Asn

225 230 235 240

Thr Arg Asp Ala Ile Ala Lys Leu Gly Thr Leu Gln Phe Thr Asp Lys

245 250 255

Gln Leu Ala Thr Ile Gly Ala Gly Val Lys Pro Ile Asp Glu Ala Ser

260 265 270

Leu Val Lys Lys Ile Val Asp Gly Val Arg Ala Leu Phe Gly Arg Ala

275 280 285

Ala Ala

290

<210> 40

<211> 185

<212> PROTEIN

<213> (Phi)AB2

<400> 40

Met Ile Leu Thr Lys Asp Gly Phe Ser Ile Ile Arg Asn Glu Leu Phe

1 5 10 15

Gly Gly Lys Leu Asp Gln Thr Gln Val Asp Ala Ile Asn Phe Ile Val

20 25 30

Ala Lys Ala Thr Glu Ser Gly Leu Thr Tyr Pro Glu Ala Ala Tyr Leu

35 40 45

Leu Ala Thr Ile Tyr His Glu Thr Gly Leu Pro Ser Gly Tyr Arg Thr

50 55 60

Met Gln Pro Ile Lys Glu Ala Gly Ser Asp Ser Tyr Leu Arg Ser Lys

65 70 75 80

Lys Tyr Tyr Pro Tyr Ile Gly Tyr Gly Tyr Val Gln Leu Thr Trp Lys

85 90 95

Glu Asn Tyr Glu Arg Ile Gly Lys Leu Ile Gly Val Asp Leu Ile Lys

100 105 110

Asn Pro Glu Lys Ala Leu Glu Pro Leu Ile Ala Ile Gln Ile Ala Ile

115 120 125

Lys Gly Met Leu Asn Gly Trp Phe Thr Gly Val Gly Phe Arg Arg Lys

130 135 140

Arg Pro Val Ser Lys Tyr Asn Lys Gln Gln Tyr Val Ala Ala Arg Asn

145 150 155 160

Ile Ile Asn Gly Lys Asp Lys Ala Glu Leu Ile Ala Lys Tyr Ala Ile

165 170 175

Ile Phe Glu Arg Ala Leu Arg Ser Leu

180 185

<210> 41

<211> 262

<212> PROTEIN

<213> (Phi)B4

<400> 41

Met Ala Met Ala Leu Gln Thr Leu Ile Asp Lys Ala Asn Arg Lys Leu

1 5 10 15

Asn Val Ser Gly Met Arg Lys Asp Val Ala Asp Arg Thr Arg Ala Val

20 25 30

Ile Thr Gln Met His Ala Gln Gly Ile Tyr Ile Cys Val Ala Gln Gly

35 40 45

Phe Arg Ser Phe Ala Glu Gln Asn Ala Leu Tyr Ala Gln Gly Arg Thr

50 55 60

Lys Pro Gly Ser Ile Val Thr Asn Ala Arg Gly Gly Gln Ser Asn His

65 70 75 80

Asn Tyr Gly Val Ala Val Asp Leu Cys Leu Tyr Thr Gln Asp Gly Ser

85 90 95

Asp Val Ile Trp Thr Val Glu Gly Asn Phe Arg Lys Val Ile Ala Ala

100 105 110

Met Lys Ala Gln Gly Phe Lys Trp Gly Gly Asp Trp Val Ser Phe Lys

115 120 125

Asp Tyr Pro His Phe Glu Leu Tyr Asp Val Val Gly Gly Gln Lys Pro

130 135 140

Pro Ala Asp Asn Gly Gly Ala Val Asp Asn Gly Gly Gly Ser Gly Ser

145 150 155 160

Thr Gly Gly Ser Gly Gly Gly Ser Thr Gly Gly Gly Ser Thr Gly Gly

165 170 175

Gly Tyr Asp Ser Ser Trp Phe Thr Lys Glu Thr Gly Thr Phe Val Thr

180 185 190

Asn Thr Ser Ile Lys Leu Arg Thr Ala Pro Phe Thr Ser Ala Asp Val

195 200 205

Ile Ala Thr Leu Pro Ala Gly Ser Pro Val Asn Tyr Asn Gly Phe Gly

210 215 220

Ile Glu Tyr Asp Gly Tyr Val Trp Ile Arg Gln Pro Arg Ser Asn Gly

225 230 235 240

Tyr Gly Tyr Leu Ala Thr Gly Glu Ser Lys Gly Gly Lys Arg Gln Asn

245 250 255

Tyr Trp Gly Thr Phe Lys

260

<210> 42

<211> 274

<212> PROTEIN

<213> (Phi)CTP1

<400> 42

Met Lys Lys Ile Ala Asp Ile Ser Asn Leu Asn Gly Asn Val Asp Val

1 5 10 15

Lys Leu Leu Phe Asn Leu Gly Tyr Ile Gly Ile Ile Ala Lys Ala Ser

20 25 30

Glu Gly Gly Thr Phe Val Asp Lys Tyr Tyr Lys Gln Asn Tyr Thr Asn

35 40 45

Thr Lys Ala Gln Gly Lys Ile Thr Gly Ala Tyr His Phe Ala Asn Phe

50 55 60

Ser Thr Ile Ala Lys Ala Gln Gln Glu Ala Asn Phe Phe Leu Asn Cys

65 70 75 80

Ile Ala Gly Thr Thr Pro Asp Phe Val Val Leu Asp Leu Glu Gln Gln

85 90 95

Cys Thr Gly Asp Ile Thr Asp Ala Cys Leu Ala Phe Leu Asn Ile Val

100 105 110

Ala Lys Lys Phe Lys Cys Val Val Tyr Cys Asn Ser Ser Phe Ile Lys

115 120 125

Glu His Leu Asn Ser Lys Ile Cys Ala Tyr Pro Leu Trp Ile Ala Asn

130 135 140

Tyr Gly Val Ala Thr Pro Ala Phe Thr Leu Trp Thr Lys Tyr Ala Met

145 150 155 160

Trp Gln Phe Thr Glu Lys Gly Gln Val Ser Gly Ile Ser Gly Tyr Ile

165 170 175

Asp Phe Ser Tyr Ile Thr Asp Glu Phe Ile Lys Tyr Ile Lys Gly Glu

180 185 190

Asp Glu Val Glu Asn Leu Val Val Tyr Asn Asp Gly Ala Asp Gln Arg

195 200 205

Ala Ala Glu Tyr Leu Ala Asp Arg Leu Ala Cys Pro Thr Ile Asn Asn

210 215 220

Ala Arg Lys Phe Asp Tyr Ser Asn Val Lys Asn Val Tyr Ala Val Gly

225 230 235 240

Gly Asn Lys Glu Gln Tyr Thr Ser Tyr Leu Thr Thr Leu Ile Ala Gly

245 250 255

Ser Thr Arg Tyr Thr Thr Met Gln Ala Val Leu Asp Tyr Ile Lys Asn

260 265 270

Leu Lys

<210> 43

<211> 628

<212> PROTEIN

<213> Staphylococcus virus 187

<400> 43

Met Ala Leu Pro Lys Thr Gly Lys Pro Thr Ala Lys Gln Val Val Asp

1 5 10 15

Trp Ala Ile Asn Leu Ile Gly Ser Gly Val Asp Val Asp Gly Tyr Tyr

20 25 30

Gly Arg Gln Cys Trp Asp Leu Pro Asn Tyr Ile Phe Asn Arg Tyr Trp

35 40 45

Asn Phe Lys Thr Pro Gly Asn Ala Arg Asp Met Ala Trp Tyr Arg Tyr

50 55 60

Pro Glu Gly Phe Lys Val Phe Arg Asn Thr Ser Asp Phe Val Pro Lys

65 70 75 80

Pro Gly Asp Ile Ala Val Trp Thr Gly Gly Asn Tyr Asn Trp Asn Thr

85 90 95

Trp Gly His Thr Gly Ile Val Val Gly Pro Ser Thr Lys Ser Tyr Phe

100 105 110

Tyr Ser Val Asp Gln Asn Trp Asn Asn Ser Asn Ser Tyr Val Gly Ser

115 120 125

Pro Ala Ala Lys Ile Lys His Ser Tyr Phe Gly Val Thr His Phe Val

130 135 140

Arg Pro Ala Tyr Lys Ala Glu Pro Lys Pro Thr Pro Pro Ala Gln Asn

145 150 155 160

Asn Pro Ala Pro Lys Asp Pro Glu Pro Ser Lys Lys Pro Glu Ser Asn

165 170 175

Lys Pro Ile Tyr Lys Val Val Thr Lys Ile Leu Phe Thr Thr Ala His

180 185 190

Ile Glu His Val Lys Ala Asn Arg Phe Val His Tyr Ile Thr Lys Ser

195 200 205

Asp Asn His Asn Asn Lys Pro Asn Lys Ile Val Ile Lys Asn Thr Asn

210 215 220

Thr Ala Leu Ser Thr Ile Asp Val Tyr Arg Tyr Arg Asp Glu Leu Asp

225 230 235 240

Lys Asp Glu Ile Pro His Phe Phe Val Asp Arg Leu Asn Val Trp Ala

245 250 255

Cys Arg Pro Ile Glu Asp Ser Ile Asn Gly Tyr His Asp Ser Val Val

260 265 270

Leu Ser Ile Thr Glu Thr Arg Thr Ala Leu Ser Asp Asn Phe Lys Met

275 280 285

Asn Glu Ile Glu Cys Leu Ser Leu Ala Glu Ser Ile Leu Lys Ala Asn

290 295 300

Asn Lys Lys Met Ser Ala Ser Asn Ile Ile Val Asp Asn Lys Ala Trp

305 310 315 320

Arg Thr Phe Lys Leu His Thr Gly Lys Asp Ser Leu Lys Ser Ser Ser

325 330 335

Phe Thr Ser Lys Asp Tyr Gln Lys Ala Val Asn Glu Leu Ile Lys Leu

340 345 350

Phe Asn Asp Lys Asp Lys Leu Leu Asn Asn Lys Pro Lys Asp Val Val

355 360 365

Glu Arg Ile Arg Ile Arg Thr Ile Val Lys Glu Asn Thr Lys Phe Val

370 375 380

Pro Ser Glu Leu Lys Pro Arg Asn Asn Ile Arg Asp Lys Gln Asp Ser

385 390 395 400

Lys Ile Asp Arg Val Ile Asn Asn Tyr Thr Leu Lys Gln Ala Leu Asn

405 410 415

Ile Gln Tyr Lys Leu Asn Pro Lys Pro Gln Thr Ser Asn Gly Val Ser

420 425 430

Trp Tyr Asn Ala Ser Val Asn Gln Ile Lys Ser Ala Met Asp Thr Thr

435 440 445

Lys Ile Phe Asn Asn Asn Val Gln Val Tyr Gln Phe Leu Lys Leu Asn

450 455 460

Gln Tyr Gln Gly Ile Pro Val Asp Lys Leu Asn Lys Leu Leu Val Gly

465 470 475 480

Lys Gly Thr Leu Ala Asn Gln Gly His Ala Phe Ala Asp Gly Cys Lys

485 490 495

Lys Tyr Asn Ile Asn Glu Ile Tyr Leu Ile Ala His Arg Phe Leu Glu

500 505 510

Ser Ala Asn Gly Thr Ser Phe Phe Ala Ser Gly Lys Thr Gly Val Tyr

515 520 525

Asn Tyr Phe Gly Ile Gly Ala Phe Asp Asn Asn Pro Asn Asn Ala Met

530 535 540

Ala Phe Ala Arg Ser His Gly Trp Thr Ser Pro Thr Lys Ala Ile Ile

545 550 555 560

Gly Gly Ala Glu Phe Val Gly Lys Gly Tyr Phe Asn Val Gly Gln Asn

565 570 575

Thr Leu Tyr Arg Met Arg Trp Asn Pro Gln Lys Pro Gly Thr His Gln

580 585 590

Tyr Ala Thr Asp Ile Ser Trp Ala Lys Val Gln Ala Gln Met Ile Ser

595 600 605

Ala Met Tyr Lys Glu Ile Gly Leu Thr Gly Asp Tyr Phe Ile Tyr Asp

610 615 620

Gln Tyr Lys Lys

625

<210> 44

<211> 291

<212> PROTEIN

<213> (Phi)P35

<400> 44

Met Ala Arg Lys Phe Thr Lys Ala Glu Leu Val Ala Lys Ala Glu Lys

1 5 10 15

Lys Val Gly Gly Leu Lys Pro Asp Val Lys Lys Ala Val Leu Ser Ala

20 25 30

Val Lys Glu Ala Tyr Asp Arg Tyr Gly Ile Gly Ile Ile Val Ser Gln

35 40 45

Gly Tyr Arg Ser Ile Ala Glu Gln Asn Gly Leu Tyr Ala Gln Gly Arg

50 55 60

Thr Lys Pro Gly Asn Ile Val Thr Asn Ala Lys Gly Gly Gln Ser Asn

65 70 75 80

His Asn Phe Gly Val Ala Val Asp Phe Ala Ile Asp Leu Ile Asp Asp

85 90 95

Gly Lys Ile Asp Ser Trp Gln Pro Ser Ala Thr Ile Val Asn Met Met

100 105 110

Lys Arg Arg Gly Phe Lys Trp Gly Gly Asp Trp Lys Ser Phe Thr Asp

115 120 125

Leu Pro His Phe Glu Ala Cys Asp Trp Tyr Arg Gly Glu Arg Lys Tyr

130 135 140

Lys Val Asp Thr Ser Glu Trp Lys Lys Lys Glu Asn Ile Asn Ile Val

145 150 155 160

Ile Lys Asp Val Gly Tyr Phe Gln Asp Lys Pro Gln Phe Leu Asn Ser

165 170 175

Lys Ser Val Arg Gln Trp Lys His Gly Thr Lys Val Lys Leu Thr Lys

180 185 190

His Asn Ser His Trp Tyr Thr Gly Val Val Lys Asp Gly Asn Lys Ser

195 200 205

Val Arg Gly Tyr Ile Tyr His Ser Met Ala Lys Val Thr Ser Lys Asn

210 215 220

Ser Asp Gly Ser Val Asn Ala Thr Ile Asn Ala His Ala Phe Cys Trp

225 230 235 240

Asp Asn Lys Lys Leu Asn Gly Gly Asp Phe Ile Asn Leu Lys Arg Gly

245 250 255

Phe Lys Gly Ile Thr His Pro Ala Ser Asp Gly Phe Tyr Pro Leu Tyr

260 265 270

Phe Ala Ser Arg Lys Lys Thr Phe Tyr Ile Pro Arg Tyr Met Phe Asp

275 280 285

Ile Lys Lys

290

<210> 45

<211> 342

<212> PROTEIN

<213> (Phi)CP-7

<400> 45

Met Val Lys Lys Asn Asp Leu Phe Val Asp Val Ala Ser His Gln Gly

1 5 10 15

Tyr Asp Ile Ser Gly Ile Leu Glu Glu Ala Gly Thr Thr Asn Thr Ile

20 25 30

Ile Lys Val Ser Glu Ser Thr Ser Tyr Leu Asn Pro Cys Leu Ser Ala

35 40 45

Gln Val Ser Gln Ser Asn Pro Ile Gly Phe Tyr His Phe Ala Trp Phe

50 55 60

Gly Gly Asn Glu Glu Glu Ala Glu Ala Glu Ala Arg Tyr Phe Leu Asp

65 70 75 80

Asn Val Pro Thr Gln Val Lys Tyr Leu Val Leu Asp Tyr Glu Asp His

85 90 95

Ala Ser Ala Ser Val Gln Arg Asn Thr Thr Ala Cys Leu Arg Phe Met

100 105 110

Gln Ile Ile Ala Glu Ala Gly Tyr Thr Pro Ile Tyr Tyr Ser Tyr Lys

115 120 125

Pro Phe Thr Leu Asp Asn Val Asp Tyr Gln Gln Ile Leu Ala Gln Phe

130 135 140

Pro Asn Ser Leu Trp Ile Ala Gly Tyr Gly Leu Asn Asp Gly Thr Ala

145 150 155 160

Asn Phe Glu Tyr Phe Pro Ser Met Asp Gly Ile Arg Trp Trp Gln Tyr

165 170 175

Ser Ser Asn Pro Phe Asp Lys Asn Ile Val Leu Leu Asp Asp Glu Lys

180 185 190

Glu Asp Asn Ile Asn Asn Glu Asn Thr Leu Lys Ser Leu Thr Thr Val

195 200 205

Ala Asn Glu Val Ile Gln Gly Leu Trp Gly Asn Gly Gln Glu Arg Tyr

210 215 220

Asp Ser Leu Ala Asn Ala Gly Tyr Asp Pro Gln Ala Val Gln Asp Lys

225 230 235 240

Val Asn Glu Ile Leu Asn Ala Arg Glu Ile Ala Asp Leu Thr Thr Val

245 250 255

Ala Asn Glu Val Ile Gln Gly Leu Trp Gly Asn Gly Gln Glu Arg Tyr

260 265 270

Asp Ser Leu Ala Asn Ala Gly Tyr Asp Pro Gln Ala Val Gln Asp Lys

275 280 285

Val Asn Glu Ile Leu Asn Ala Arg Glu Ile Ala Asp Leu Thr Thr Val

290 295 300

Ala Asn Glu Val Ile Gln Gly Leu Trp Gly Asn Gly Gln Glu Arg Tyr

305 310 315 320

Asp Ser Leu Ala Asn Ala Gly Tyr Asp Pro Gln Ala Val Gln Asp Lys

325 330 335

Val Asn Glu Leu Leu Ser

340

<210> 46

<211> 328

<212> PROTEIN

<213> (Phi)EFAP-1

<400> 46

Met Lys Leu Lys Gly Ile Leu Leu Ser Val Val Thr Thr Phe Gly Leu

1 5 10 15

Leu Phe Gly Ala Thr Asn Val Gln Ala Tyr Glu Val Asn Asn Glu Phe

20 25 30

Asn Leu Gln Pro Trp Glu Gly Ser Gln Gln Leu Ala Tyr Pro Asn Lys

35 40 45

Ile Ile Leu His Glu Thr Ala Asn Pro Arg Ala Thr Gly Arg Asn Glu

50 55 60

Ala Thr Tyr Met Lys Asn Asn Trp Phe Asn Ala His Thr Thr Ala Ile

65 70 75 80

Val Gly Asp Gly Gly Ile Val Tyr Lys Val Ala Pro Glu Gly Asn Val

85 90 95

Ser Trp Gly Ala Gly Asn Ala Asn Pro Tyr Ala Pro Val Gln Ile Glu

100 105 110

Leu Gln His Thr Asn Asp Pro Glu Leu Phe Lys Ala Asn Tyr Lys Ala

115 120 125

Tyr Val Asp Tyr Thr Arg Asp Met Gly Lys Lys Phe Gly Ile Pro Met

130 135 140

Thr Leu Asp Gln Gly Gly Ser Leu Trp Glu Lys Gly Val Val Ser His

145 150 155 160

Gln Trp Val Thr Asp Phe Val Trp Gly Asp His Thr Asp Pro Tyr Gly

165 170 175

Tyr Leu Ala Lys Met Gly Ile Ser Lys Ala Gln Leu Ala His Asp Leu

180 185 190

Ala Asn Gly Val Ser Gly Asn Thr Ala Thr Pro Thr Pro Lys Pro Asp

195 200 205

Lys Pro Lys Pro Thr Gln Pro Ser Lys Pro Ser Asn Lys Lys Arg Phe

210 215 220

Asn Tyr Arg Val Asp Gly Leu Glu Tyr Val Asn Gly Met Trp Gln Ile

225 230 235 240

Tyr Asn Glu His Leu Gly Lys Ile Asp Phe Asn Trp Thr Glu Asn Gly

245 250 255

Ile Pro Val Glu Val Val Asp Lys Val Asn Pro Ala Thr Gly Gln Pro

260 265 270

Thr Lys Asp Gln Val Leu Lys Val Gly Asp Tyr Phe Asn Phe Gln Glu

275 280 285

Asn Ser Thr Gly Val Val Gln Glu Gln Thr Pro Tyr Met Gly Tyr Thr

290 295 300

Leu Ser His Val Gln Leu Pro Asn Glu Phe Ile Trp Leu Phe Thr Asp

305 310 315 320

Ser Lys Gln Ala Leu Met Tyr Gln

325

<210> 47

<211> 48

<212> PROTEIN

<213> Homo sapiens

<400> 47

Ser Ser Leu Leu Glu Lys Gly Leu Asp Gly Ala Lys Lys Ala Val Gly

1 5 10 15

Gly Leu Gly Lys Leu Gly Lys Asp Ala Val Glu Asp Leu Glu Ser Val

20 25 30

Gly Lys Gly Ala Val His Asp Val Lys Asp Val Leu Asp Ser Val Leu

35 40 45

<210> 48

<211> 37

<212> PROTEIN

<213> Hyalophora cecropia

<400> 48

Lys Trp Lys Leu Phe Lys Lys Ile Glu Lys Val Gly Gln Asn Ile Arg

1 5 10 15

Asp Gly Ile Ile Lys Ala Gly Pro Ala Val Ala Val Val Gly Gln Ala

20 25 30

Thr Gln Ile Ala Lys

35

<210> 49

<211> 62

<212> PROTEIN

<213> Drosophila teissieri

<400> 49

Met Lys Tyr Phe Ser Val Leu Val Val Leu Thr Leu Ile Leu Ala Ile

1 5 10 15

Val Asp Gln Ser Asp Ala Phe Ile Asn Leu Leu Asp Lys Val Glu Asp

20 25 30

Ala Leu His Thr Gly Ala Gln Ala Gly Phe Lys Leu Ile Arg Pro Val

35 40 45

Glu Arg Gly Ala Thr Pro Lys Lys Ser Glu Lys Pro Glu Lys

50 55 60

<210> 50

<211> 66

<212> PROTEIN

<213> Bombyx mori

<400> 50

Met Asn Ile Leu Lys Phe Phe Phe Val Phe Ile Val Ala Met Ser Leu

1 5 10 15

Val Ser Cys Ser Thr Ala Ala Pro Ala Lys Ile Pro Ile Lys Ala Ile

20 25 30

Lys Thr Val Gly Lys Ala Val Gly Lys Gly Leu Arg Ala Ile Asn Ile

35 40 45

Ala Ser Thr Ala Asn Asp Val Phe Asn Phe Leu Lys Pro Lys Lys Arg

50 55 60

Lys His

65

<210> 51

<211> 71

<212> PROTEIN

<213> Ceratitis capitata

<400> 51

Met Ala Asn Leu Lys Ala Val Phe Leu Ile Cys Ile Val Ala Phe Ile

1 5 10 15

Ala Leu Gln Cys Val Val Ala Glu Pro Ala Ala Glu Asp Ser Val Val

20 25 30

Val Lys Arg Ser Ile Gly Ser Ala Leu Lys Lys Ala Leu Pro Val Ala

35 40 45

Lys Lys Ile Gly Lys Ile Ala Leu Pro Ile Ala Lys Ala Ala Leu Pro

50 55 60

Val Ala Ala Gly Leu Val Gly

65 70

<210> 52

<211> 53

<212> PROTEIN

<213> Apis mellifera

<400> 52

Met Lys Val Val Ile Phe Ile Phe Ala Leu Leu Ala Thr Ile Cys Ala

1 5 10 15

Ala Phe Ala Tyr Val Pro Leu Pro Asn Val Pro Gln Pro Gly Arg Arg

20 25 30

Pro Phe Pro Thr Phe Pro Gly Gln Gly Pro Phe Asn Pro Lys Ile Lys

35 40 45

Trp Pro Gln Gly Tyr

50

<210> 53

<211> 283

<212> PROTEIN

<213> Apis mellifera

<400> 53

Lys Asn Phe Ala Leu Ala Ile Leu Val Val Thr Phe Val Val Ala Val

1 5 10 15

Phe Gly Asn Thr Asn Leu Asp Pro Pro Thr Arg Pro Thr Arg Leu Arg

20 25 30

Arg Glu Ala Lys Pro Glu Ala Glu Pro Gly Asn Asn Arg Pro Val Tyr

35 40 45

Ile Pro Gln Pro Arg Pro Pro His Pro Arg Leu Arg Arg Glu Ala Glu

50 55 60

Pro Glu Ala Glu Pro Gly Asn Asn Arg Pro Val Tyr Ile Pro Gln Pro

65 70 75 80

Arg Pro Pro His Pro Arg Leu Arg Arg Glu Ala Glu Leu Glu Ala Glu

85 90 95

Pro Gly Asn Asn Arg Pro Val Tyr Ile Ser Gln Pro Arg Pro Pro His

100 105 110

Pro Arg Leu Arg Arg Glu Ala Glu Pro Glu Ala Glu Pro Gly Asn Asn

115 120 125

Arg Pro Val Tyr Ile Pro Gln Pro Arg Pro Pro His Pro Arg Leu Arg

130 135 140

Arg Glu Ala Glu Leu Glu Ala Glu Pro Gly Asn Asn Arg Pro Val Tyr

145 150 155 160

Ile Ser Gln Pro Arg Pro Pro His Pro Arg Leu Arg Arg Glu Ala Glu

165 170 175

Pro Glu Ala Glu Pro Gly Asn Asn Arg Pro Val Tyr Ile Pro Gln Pro

180 185 190

Arg Pro Pro His Pro Arg Leu Arg Arg Glu Ala Glu Pro Glu Ala Glu

195 200 205

Pro Gly Asn Asn Arg Pro Val Tyr Ile Pro Gln Pro Arg Pro Pro His

210 215 220

Pro Arg Leu Arg Arg Glu Ala Glu Pro Glu Ala Glu Pro Gly Asn Asn

225 230 235 240

Arg Pro Val Tyr Ile Pro Gln Pro Arg Pro Pro His Pro Arg Leu Arg

245 250 255

Arg Glu Ala Lys Pro Glu Ala Lys Pro Gly Asn Asn Arg Pro Val Tyr

260 265 270

Ile Pro Gln Pro Arg Pro Pro His Pro Arg Ile

275 280

<210> 54

<211> 228

<212> PROTEIN

<213> Sus scrofa

<400> 54

Met Glu Thr Gln Arg Ala Ser Leu Cys Leu Gly Arg Trp Ser Leu Trp

1 5 10 15

Leu Leu Leu Leu Ala Leu Val Val Pro Ser Ala Ser Ala Gln Ala Leu

20 25 30

Ser Tyr Arg Glu Ala Val Leu Arg Ala Val Asp Arg Leu Asn Glu Gln

35 40 45

Ser Ser Glu Ala Asn Leu Tyr Arg Leu Leu Glu Leu Asp Gln Pro Pro

50 55 60

Lys Ala Asp Glu Asp Pro Gly Thr Pro Lys Pro Val Ser Phe Thr Val

65 70 75 80

Lys Glu Thr Val Cys Pro Arg Pro Thr Arg Arg Pro Pro Glu Leu Cys

85 90 95

Asp Phe Lys Glu Asn Gly Arg Val Lys Gln Cys Val Gly Thr Val Thr

100 105 110

Leu Asp Gln Ile Lys Asp Pro Leu Asp Ile Thr Cys Asn Glu Gly Val

115 120 125

Arg Arg Phe Pro Trp Trp Trp Pro Phe Leu Arg Arg Pro Arg Leu Arg

130 135 140

Arg Gln Ala Phe Pro Pro Pro Asn Val Pro Gly Pro Arg Phe Pro Pro

145 150 155 160

Pro Asn Val Pro Gly Pro Arg Phe Pro Pro Pro Asn Phe Pro Gly Pro

165 170 175

Arg Phe Pro Pro Pro Asn Phe Pro Gly Pro Arg Phe Pro Pro Pro Asn

180 185 190

Phe Pro Gly Pro Pro Phe Pro Pro Pro Ile Phe Pro Gly Pro Trp Phe

195 200 205

Pro Pro Pro Pro Pro Phe Arg Pro Pro Pro Phe Gly Pro Pro Arg Phe

210 215 220

Pro Gly Arg Arg

225

<210> 55

<211> 144

<212> PROTEIN

<213> Bos taurus

<400> 55

Met Gln Thr Gln Arg Ala Ser Leu Ser Leu Gly Arg Trp Ser Leu Trp

1 5 10 15

Leu Leu Leu Leu Gly Leu Val Val Pro Ser Ala Ser Ala Gln Ala Leu

20 25 30

Ser Tyr Arg Glu Ala Val Leu Arg Ala Val Asp Gln Leu Asn Glu Leu

35 40 45

Ser Ser Glu Ala Asn Leu Tyr Arg Leu Leu Glu Leu Asp Pro Pro Pro

50 55 60

Lys Asp Asn Glu Asp Leu Gly Thr Arg Lys Pro Val Ser Phe Thr Val

65 70 75 80

Lys Glu Thr Val Cys Pro Arg Thr Ile Gln Gln Pro Ala Glu Gln Cys

85 90 95

Asp Phe Lys Glu Lys Gly Arg Val Lys Gln Cys Val Gly Thr Val Thr

100 105 110

Leu Asp Pro Ser Asn Asp Gln Phe Asp Leu Asn Cys Asn Glu Leu Gln

115 120 125

Ser Val Ile Leu Pro Trp Lys Trp Pro Trp Trp Pro Trp Arg Arg Gly

130 135 140

<210> 56

<211> 149

<212> PROTEIN

<213> Sus scrofa

<400> 56

Met Glu Thr Gln Arg Ala Ser Leu Cys Leu Gly Arg Trp Ser Leu Trp

1 5 10 15

Leu Leu Leu Leu Ala Leu Val Val Pro Ser Ala Ser Ala Gln Ala Leu

20 25 30

Ser Tyr Arg Glu Ala Val Leu Arg Ala Val Asp Arg Leu Asn Glu Gln

35 40 45

Ser Ser Glu Ala Asn Leu Tyr Arg Leu Leu Glu Leu Asp Gln Pro Pro

50 55 60

Lys Ala Asp Glu Asp Pro Gly Thr Pro Lys Pro Val Ser Phe Thr Val

65 70 75 80

Lys Glu Thr Val Cys Pro Arg Pro Thr Arg Gln Pro Pro Glu Leu Cys

85 90 95

Asp Phe Lys Glu Asn Gly Arg Val Lys Gln Cys Val Gly Thr Val Thr

100 105 110

Leu Asp Gln Ile Lys Asp Pro Leu Asp Ile Thr Cys Asn Glu Val Gln

115 120 125

Gly Val Arg Gly Gly Arg Leu Cys Tyr Cys Arg Arg Arg Phe Cys Val

130 135 140

Cys Val Gly Arg Gly

145

<210> 57

<211> 17

<212> PROTEIN

<213> Tachypleus gigas

<400> 57

Lys Trp Cys Phe Arg Val Cys Tyr Arg Gly Ile Cys Tyr Arg Arg Cys

1 5 10 15

Arg

<210> 58

<211> 102

<212> PROTEIN

<213> Anopheles gambiae

<400> 58

Met Lys Cys Ala Thr Ile Val Cys Thr Ile Ala Val Val Leu Ala Ala

1 5 10 15

Thr Leu Leu Asn Gly Ser Val Gln Ala Ala Pro Gln Glu Glu Ala Ala

20 25 30

Leu Ser Gly Gly Ala Asn Leu Asn Thr Leu Leu Asp Glu Leu Pro Glu

35 40 45

Glu Thr His His Ala Ala Leu Glu Asn Tyr Arg Ala Lys Arg Ala Thr

50 55 60

Cys Asp Leu Ala Ser Gly Phe Gly Val Gly Ser Ser Leu Cys Ala Ala

65 70 75 80

His Cys Ile Ala Arg Arg Tyr Arg Gly Gly Tyr Cys Asn Ser Lys Ala

85 90 95

Val Cys Val Cys Arg Asn

100

<210> 59

<211> 70

<212> PROTEIN

<213> Drosophila melanogaster

<400> 59

Met Met Gln Ile Lys Tyr Leu Phe Ala Leu Phe Ala Val Leu Met Leu

1 5 10 15

Val Val Leu Gly Ala Asn Glu Ala Asp Ala Asp Cys Leu Ser Gly Arg

20 25 30

Tyr Lys Gly Pro Cys Ala Val Trp Asp Asn Glu Thr Cys Arg Arg Val

35 40 45

Cys Lys Glu Glu Gly Arg Ser Ser Gly His Cys Ser Pro Ser Leu Lys

50 55 60

Cys Trp Cys Glu Gly Cys

65 70

<210> 60

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 60

Met Thr Lys Ile Val Val Phe Ile Tyr Val Val Ile Leu Leu Leu Thr

1 5 10 15

Ile Phe His Val Ser Ala Lys Lys Lys Arg Tyr Ile Glu Cys Glu Thr

20 25 30

His Glu Asp Cys Ser Gln Val Phe Met Pro Pro Phe Val Met Arg Cys

35 40 45

Val Ile His Glu Cys Lys Ile Phe Asn Gly Glu His Leu Arg Tyr

50 55 60

<210> 61

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 61

Met Ala Lys Ile Met Lys Phe Val Tyr Asn Met Ile Pro Phe Leu Ser

1 5 10 15

Ile Phe Ile Ile Thr Leu Gln Val Asn Val Val Val Cys Glu Ile Asp

20 25 30

Ala Asp Cys Pro Gln Ile Cys Met Pro Pro Tyr Glu Val Arg Cys Val

35 40 45

Asn His Arg Cys Gly Trp Val Asn Thr Asp Asp Ser Leu Phe Leu Thr

50 55 60

Gln Glu Phe Thr Arg Ser Lys Gln Tyr Ile Ile Ser

65 70 75

<210> 62

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 62

Met Tyr Lys Val Val Glu Ser Ile Phe Ile Arg Tyr Met His Arg Lys

1 5 10 15

Pro Asn Met Thr Lys Phe Phe Lys Phe Val Tyr Thr Met Phe Ile Leu

20 25 30

Ile Ser Leu Phe Leu Val Val Thr Asn Ala Asn Ala His Asn Cys Thr

35 40 45

Asp Ile Ser Asp Cys Ser Ser Asn His Cys Ser Tyr Glu Gly Val Ser

50 55 60

Leu Cys Met Asn Gly Gln Cys Ile Cys Ile Tyr Glu

65 70 75

<210> 63

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 63

Met Val Glu Thr Leu Arg Leu Phe Tyr Ile Met Ile Leu Phe Val Ser

1 5 10 15

Leu Cys Leu Val Val Val Asp Gly Glu Ser Lys Leu Glu Gln Thr Cys

20 25 30

Ser Glu Asp Phe Glu Cys Tyr Ile Lys Asn Pro His Val Pro Phe Gly

35 40 45

His Leu Arg Cys Phe Glu Gly Phe Cys Gln Gln Leu Asn Gly Pro Ala

50 55 60

<210> 64

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 64

Met Ala Lys Ile Val Asn Phe Val Tyr Ser Met Ile Val Phe Leu Phe

1 5 10 15

Leu Phe Leu Val Ala Thr Lys Ala Ala Arg Gly Tyr Leu Cys Val Thr

20 25 30

Asp Ser His Cys Pro Pro His Met Cys Pro Pro Gly Met Glu Pro Arg

35 40 45

Cys Val Arg Arg Met Cys Lys Cys Leu Pro Ile Gly Trp Arg Lys Tyr

50 55 60

Phe Val Pro

65

<210> 65

<211> 96

<212> PROTEIN

<213> Medicago truncatula

<400> 65

Met Gln Ile Gly Lys Asn Met Val Glu Thr Pro Lys Leu Asp Tyr Val

1 5 10 15

Ile Ile Phe Phe Phe Leu Tyr Phe Phe Phe Arg Gln Met Ile Ile Leu

20 25 30

Arg Leu Asn Thr Thr Phe Arg Pro Leu Asn Phe Lys Met Leu Arg Phe

35 40 45

Trp Gly Gln Asn Arg Asn Ile Met Lys His Arg Gly Gln Lys Val His

50 55 60

Phe Ser Leu Ile Leu Ser Asp Cys Lys Thr Asn Lys Asp Cys Pro Lys

65 70 75 80

Leu Arg Arg Ala Asn Val Arg Cys Arg Lys Ser Tyr Cys Val Pro Ile

85 90 95

<210> 66

<211> 65

<212> PROTEIN

<213> Medicago truncatula

<400> 66

Met Leu Arg Leu Tyr Leu Val Ser Tyr Phe Leu Leu Lys Arg Thr Leu

1 5 10 15

Leu Val Ser Tyr Phe Ser Tyr Phe Ser Thr Tyr Ile Ile Glu Cys Lys

20 25 30

Thr Asp Asn Asp Cys Pro Ile Ser Gln Leu Lys Ile Tyr Ala Trp Lys

35 40 45

Cys Val Lys Asn Gly Cys His Leu Phe Asp Val Ile Pro Met Met Tyr

50 55 60

Glu

65

<210> 67

<211> 79

<212> PROTEIN

<213> Medicago truncatula

<400> 67

Met Ala Glu Ile Leu Lys Phe Val Tyr Ile Val Ile Leu Phe Val Ser

1 5 10 15

Leu Leu Leu Ile Val Val Ala Ser Glu Arg Glu Cys Val Thr Asp Asp

20 25 30

Asp Cys Glu Lys Leu Tyr Pro Thr Asn Glu Tyr Arg Met Met Cys Asp

35 40 45

Ser Gly Tyr Cys Met Asn Leu Leu Asn Gly Lys Ile Ile Tyr Leu Leu

50 55 60

Cys Leu Lys Lys Lys Lys Phe Leu Ile Ile Ile Ser Val Leu Leu

65 70 75

<210> 68

<211> 95

<212> PROTEIN

<213> Medicago truncatula

<400> 68

Met Ala Glu Ile Ile Lys Phe Val Tyr Ile Met Ile Leu Cys Val Ser

1 5 10 15

Leu Leu Leu Ile Glu Val Ala Gly Glu Glu Cys Val Thr Asp Ala Asp

20 25 30

Cys Asp Lys Leu Tyr Pro Asp Ile Arg Lys Pro Leu Met Cys Ser Ile

35 40 45

Gly Glu Cys Tyr Ser Leu Tyr Lys Gly Lys Phe Ser Leu Ser Ile Ile

50 55 60

Ser Lys Thr Ser Phe Ser Leu Met Val Tyr Asn Val Val Thr Leu Val

65 70 75 80

Ile Cys Leu Arg Leu Ala Tyr Ile Ser Leu Leu Leu Lys Phe Leu

85 90 95

<210> 69

<211> 100

<212> PROTEIN

<213> Medicago truncatula

<400> 69

Met Ala Glu Ile Leu Lys Asp Phe Tyr Ala Met Asn Leu Phe Ile Phe

1 5 10 15

Leu Ile Ile Leu Pro Ala Lys Ile Arg Gly Glu Thr Leu Ser Leu Thr

20 25 30

His Pro Lys Cys His His Ile Met Leu Pro Ser Leu Phe Ile Thr Glu

35 40 45

Val Phe Gln Arg Val Thr Asp Asp Gly Cys Pro Lys Pro Val Asn His

50 55 60

Leu Arg Val Val Lys Cys Ile Glu His Ile Cys Glu Tyr Gly Tyr Asn

65 70 75 80

Tyr Arg Pro Asp Phe Ala Ser Gln Ile Pro Glu Ser Thr Lys Met Pro

85 90 95

Arg Lys Arg Glu

100

<210> 70

<211> 78

<212> PROTEIN

<213> Medicago truncatula

<400> 70

Met Val Glu Ile Leu Lys Asn Phe Tyr Ala Met Asn Leu Phe Ile Phe

1 5 10 15

Leu Ile Ile Leu Ala Val Lys Ile Arg Gly Ala His Phe Pro Cys Val

20 25 30

Thr Asp Asp Asp Cys Pro Lys Pro Val Asn Lys Leu Arg Val Ile Lys

35 40 45

Cys Ile Asp His Ile Cys Gln Tyr Ala Arg Asn Leu Pro Asp Phe Ala

50 55 60

Ser Glu Ile Ser Glu Ser Thr Lys Met Pro Cys Lys Gly Glu

65 70 75

<210> 71

<211> 72

<212> PROTEIN

<213> Medicago truncatula

<400> 71

Met Phe His Ala Gln Ala Glu Asn Met Ala Lys Val Ser Asn Phe Val

1 5 10 15

Cys Ile Met Ile Leu Phe Leu Ala Leu Phe Phe Ile Thr Met Asn Asp

20 25 30

Ala Ala Arg Phe Glu Cys Arg Glu Asp Ser His Cys Val Thr Arg Ile

35 40 45

Lys Cys Val Leu Pro Arg Lys Pro Glu Cys Arg Asn Tyr Ala Cys Gly

50 55 60

Cys Tyr Asp Ser Asn Lys Tyr Arg

65 70

<210> 72

<211> 78

<212> PROTEIN

<213> Medicago truncatula

<400> 72

Met Gln Met Arg Gln Asn Met Ala Thr Ile Leu Asn Phe Val Phe Val

1 5 10 15

Ile Ile Leu Phe Ile Ser Leu Leu Leu Val Val Thr Lys Gly Tyr Arg

20 25 30

Glu Pro Phe Ser Ser Phe Thr Glu Gly Pro Thr Cys Lys Glu Asp Ile

35 40 45

Asp Cys Pro Ser Ile Ser Cys Val Asn Pro Gln Val Pro Lys Cys Ile

50 55 60

Met Phe Glu Cys His Cys Lys Tyr Ile Pro Thr Thr Leu Lys

65 70 75

<210> 73

<211> 71

<212> PROTEIN

<213> Medicago truncatula

<400> 73

Met Ala Thr Ile Leu Met Tyr Val Tyr Ile Thr Ile Leu Phe Ile Ser

1 5 10 15

Ile Leu Thr Val Leu Thr Glu Gly Leu Tyr Glu Pro Leu Tyr Asn Phe

20 25 30

Arg Arg Asp Pro Asp Cys Arg Arg Asn Ile Asp Cys Pro Ser Tyr Leu

35 40 45

Cys Val Ala Pro Lys Val Pro Arg Cys Ile Met Phe Glu Cys His Cys

50 55 60

Lys Asp Ile Pro Ser Asp His

65 70

<210> 74

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 74

Met Thr Thr Ser Leu Lys Phe Val Tyr Val Ala Ile Leu Phe Leu Ser

1 5 10 15

Leu Leu Leu Val Val Met Gly Gly Ile Arg Arg Phe Glu Cys Arg Gln

20 25 30

Asp Ser Asp Cys Pro Ser Tyr Phe Cys Glu Lys Leu Thr Val Pro Lys

35 40 45

Cys Phe Trp Ser Lys Cys Tyr Cys Lys

50 55

<210> 75

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 75

Met Thr Thr Ser Leu Lys Phe Val Tyr Val Ala Ile Leu Phe Leu Ser

1 5 10 15

Leu Leu Leu Val Val Met Gly Gly Ile Arg Lys Lys Glu Cys Arg Gln

20 25 30

Asp Ser Asp Cys Pro Ser Tyr Phe Cys Glu Lys Leu Thr Ile Ala Lys

35 40 45

Cys Ile His Ser Thr Cys Leu Cys Lys

50 55

<210> 76

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 76

Met Gln Ile Gly Lys Asn Met Val Glu Thr Pro Lys Leu Val Tyr Phe

1 5 10 15

Ile Ile Leu Phe Leu Ser Ile Phe Leu Cys Ile Thr Val Ser Asn Ser

20 25 30

Ser Phe Ser Gln Ile Phe Asn Ser Ala Cys Lys Thr Asp Lys Asp Cys

35 40 45

Pro Lys Phe Gly Arg Val Asn Val Arg Cys Arg Lys Gly Asn Cys Val

50 55 60

Pro Ile

65

<210> 77

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 77

Met Thr Ala Ile Leu Lys Lys Phe Ile Asn Ala Val Phe Leu Phe Ile

1 5 10 15

Val Leu Phe Leu Ala Thr Thr Asn Val Glu Asp Phe Val Gly Gly Ser

20 25 30

Asn Asp Glu Cys Val Tyr Pro Asp Val Phe Gln Cys Ile Asn Asn Ile

35 40 45

Cys Lys Cys Val Ser His His Arg Thr

50 55

<210> 78

<211> 74

<212> PROTEIN

<213> Medicago truncatula

<400> 78

Met Gln Lys Arg Lys Asn Met Ala Gln Ile Ile Phe Tyr Val Tyr Ala

1 5 10 15

Leu Ile Ile Leu Phe Ser Pro Phe Leu Ala Ala Arg Leu Val Phe Val

20 25 30

Asn Pro Glu Lys Pro Cys Val Thr Asp Ala Asp Cys Asp Arg Tyr Arg

35 40 45

His Glu Ser Ala Ile Tyr Ser Asp Met Phe Cys Lys Asp Gly Tyr Cys

50 55 60

Phe Ile Asp Tyr His His Asp Pro Tyr Pro

65 70

<210> 79

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 79

Met Gln Met Arg Lys Asn Met Ala Gln Ile Leu Phe Tyr Val Tyr Ala

1 5 10 15

Leu Leu Ile Leu Phe Thr Pro Phe Leu Val Ala Arg Ile Met Val Val

20 25 30

Asn Pro Asn Asn Pro Cys Val Thr Asp Ala Asp Cys Gln Arg Tyr Arg

35 40 45

His Lys Leu Ala Thr Arg Met Ile Cys Asn Gln Gly Phe Cys Leu Met

50 55 60

Asp Phe Thr His Asp Pro Tyr Ala Pro Ser Leu Pro

65 70 75

<210> 80

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 80

Met Asn His Ile Ser Lys Phe Val Tyr Ala Leu Ile Ile Phe Leu Ser

1 5 10 15

Ile Tyr Leu Val Val Leu Asp Gly Leu Pro Ile Ser Cys Lys Asp His

20 25 30

Phe Glu Cys Arg Arg Lys Ile Asn Ile Leu Arg Cys Ile Tyr Arg Gln

35 40 45

Glu Lys Pro Met Cys Ile Asn Ser Ile Cys Thr Cys Val Lys Leu Leu

50 55 60

<210> 81

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 81

Met Gln Arg Glu Lys Asn Met Ala Lys Ile Phe Glu Phe Val Tyr Ala

1 5 10 15

Met Ile Ile Phe Ile Leu Leu Phe Leu Val Glu Lys Asn Val Val Ala

20 25 30

Tyr Leu Lys Phe Glu Cys Lys Thr Asp Asp Asp Cys Gln Lys Ser Leu

35 40 45

Leu Lys Thr Tyr Val Trp Lys Cys Val Lys Asn Glu Cys Tyr Phe Phe

50 55 60

Ala Lys Lys

65

<210> 82

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 82

Met Ala Gly Ile Ile Lys Phe Val His Val Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe His Val Val Lys Asn Asp Asp Gly Ser Phe Cys Phe Lys Asp

20 25 30

Ser Asp Cys Pro Asp Glu Met Cys Pro Ser Pro Leu Lys Glu Met Cys

35 40 45

Tyr Phe Leu Gln Cys Lys Cys Gly Val Asp Thr Ile Ala

50 55 60

<210> 83

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 83

Met Ala Asn Thr His Lys Leu Val Ser Met Ile Leu Phe Ile Phe Leu

1 5 10 15

Phe Leu Ala Ser Asn Asn Val Glu Gly Tyr Val Asn Cys Glu Thr Asp

20 25 30

Ala Asp Cys Pro Pro Ser Thr Arg Val Lys Arg Phe Lys Cys Val Lys

35 40 45

Gly Glu Cys Arg Trp Thr Arg Met Ser Tyr Ala

50 55

<210> 84

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 84

Met Gln Arg Arg Lys Lys Lys Ala Gln Val Val Met Phe Val His Asp

1 5 10 15

Leu Ile Ile Cys Ile Tyr Leu Phe Ile Val Ile Thr Thr Arg Lys Thr

20 25 30

Asp Ile Arg Cys Arg Phe Tyr Tyr Asp Cys Pro Arg Leu Glu Tyr His

35 40 45

Phe Cys Glu Cys Ile Glu Asp Phe Cys Ala Tyr Ile Arg Leu Asn

50 55 60

<210> 85

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 85

Met Ala Lys Val Tyr Met Phe Val Tyr Ala Leu Ile Ile Phe Val Ser

1 5 10 15

Pro Phe Leu Leu Ala Thr Phe Arg Thr Arg Leu Pro Cys Glu Lys Asp

20 25 30

Asp Asp Cys Pro Glu Ala Phe Leu Pro Pro Val Met Lys Cys Val Asn

35 40 45

Arg Phe Cys Gln Tyr Glu Ile Leu Glu

50 55

<210> 86

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 86

Met Ile Lys Gln Phe Ser Val Cys Tyr Ile Gln Met Arg Arg Asn Met

1 5 10 15

Thr Thr Ile Leu Lys Phe Pro Tyr Ile Met Val Ile Cys Leu Leu Leu

20 25 30

Leu His Val Ala Ala Tyr Glu Asp Phe Glu Lys Glu Ile Phe Asp Cys

35 40 45

Lys Lys Asp Gly Asp Cys Asp His Met Cys Val Thr Pro Gly Ile Pro

50 55 60

Lys Cys Thr Gly Tyr Val Cys Phe Cys Phe Glu Asn Leu

65 70 75

<210> 87

<211> 73

<212> PROTEIN

<213> Medicago truncatula

<400> 87

Met Gln Arg Ser Arg Asn Met Thr Thr Ile Phe Lys Phe Ala Tyr Ile

1 5 10 15

Met Ile Ile Cys Val Phe Leu Leu Asn Ile Ala Ala Gln Glu Ile Glu

20 25 30

Asn Gly Ile His Pro Cys Lys Lys Asn Glu Asp Cys Asn His Met Cys

35 40 45

Val Met Pro Gly Leu Pro Trp Cys His Glu Asn Asn Leu Cys Phe Cys

50 55 60

Tyr Glu Asn Ala Tyr Gly Asn Thr Arg

65 70

<210> 88

<211> 85

<212> PROTEIN

<213> Medicago truncatula

<400> 88

Met Thr Ile Ile Ile Lys Phe Val Asn Val Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe His Val Ala Lys Asn Asp Asp Asn Lys Leu Leu Leu Ser Phe

20 25 30

Ile Glu Glu Gly Phe Leu Cys Phe Lys Asp Ser Asp Cys Pro Tyr Asn

35 40 45

Met Cys Pro Ser Pro Leu Lys Glu Met Cys Tyr Phe Ile Lys Cys Val

50 55 60

Cys Gly Val Tyr Gly Pro Ile Arg Glu Arg Arg Leu Tyr Gln Ser His

65 70 75 80

Asn Pro Met Ile Gln

85

<210> 89

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 89

Met Arg Lys Asn Met Thr Lys Ile Leu Met Ile Gly Tyr Ala Leu Met

1 5 10 15

Ile Phe Ile Phe Leu Ser Ile Ala Val Ser Ile Thr Gly Asn Leu Ala

20 25 30

Arg Ala Ser Arg Lys Lys Pro Val Asp Val Ile Pro Cys Ile Tyr Asp

35 40 45

His Asp Cys Pro Arg Lys Leu Tyr Phe Leu Glu Arg Cys Val Gly Arg

50 55 60

Val Cys Lys Tyr Leu

65

<210> 90

<211> 58

<212> PROTEIN

<213> Medicago truncatula

<400> 90

Met Ala His Lys Leu Val Tyr Ala Ile Thr Leu Phe Ile Phe Leu Phe

1 5 10 15

Leu Ile Ala Asn Asn Ile Glu Asp Asp Ile Phe Cys Ile Thr Asp Asn

20 25 30

Asp Cys Pro Pro Asn Thr Leu Val Gln Arg Tyr Arg Cys Ile Asn Gly

35 40 45

Lys Cys Asn Leu Ser Phe Val Ser Tyr Gly

50 55

<210> 91

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 91

Met Asp Glu Thr Leu Lys Phe Val Tyr Ile Leu Ile Leu Phe Val Ser

1 5 10 15

Leu Cys Leu Val Val Ala Asp Gly Val Lys Asn Ile Asn Arg Glu Cys

20 25 30

Thr Gln Thr Ser Asp Cys Tyr Lys Lys Tyr Pro Phe Ile Pro Trp Gly

35 40 45

Lys Val Arg Cys Val Lys Gly Arg Cys Arg Leu Asp Met

50 55 60

<210> 92

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 92

Met Ala Lys Ile Ile Lys Phe Val Tyr Val Leu Ala Ile Phe Phe Ser

1 5 10 15

Leu Phe Leu Val Ala Lys Asn Val Asn Gly Trp Thr Cys Val Glu Asp

20 25 30

Ser Asp Cys Pro Ala Asn Ile Cys Gln Pro Pro Met Gln Arg Met Cys

35 40 45

Phe Tyr Gly Glu Cys Ala Cys Val Arg Ser Lys Phe Cys Thr

50 55 60

<210> 93

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 93

Met Val Lys Ile Ile Lys Phe Val Tyr Phe Met Thr Leu Phe Leu Ser

1 5 10 15

Met Leu Leu Val Thr Thr Lys Glu Asp Gly Ser Val Glu Cys Ile Ala

20 25 30

Asn Ile Asp Cys Pro Gln Ile Phe Met Leu Pro Phe Val Met Arg Cys

35 40 45

Ile Asn Phe Arg Cys Gln Ile Val Asn Ser Glu Asp Thr

50 55 60

<210> 94

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 94

Met Asp Glu Ile Leu Lys Phe Val Tyr Thr Leu Ile Ile Phe Phe Ser

1 5 10 15

Leu Phe Phe Ala Ala Asn Asn Val Asp Ala Asn Ile Met Asn Cys Gln

20 25 30

Ser Thr Phe Asp Cys Pro Arg Asp Met Cys Ser His Ile Arg Asp Val

35 40 45

Ile Cys Ile Phe Lys Lys Cys Lys Cys Ala Gly Gly Arg Tyr Met Pro

50 55 60

Gln Val Pro

65

<210> 95

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 95

Met Gln Arg Arg Lys Asn Met Ala Asn Asn His Met Leu Ile Tyr Ala

1 5 10 15

Met Ile Ile Cys Leu Phe Pro Tyr Leu Val Val Thr Phe Lys Thr Ala

20 25 30

Ile Thr Cys Asp Cys Asn Glu Asp Cys Leu Asn Phe Phe Thr Pro Leu

35 40 45

Asp Asn Leu Lys Cys Ile Asp Asn Val Cys Glu Val Phe Met

50 55 60

<210> 96

<211> 65

<212> PROTEIN

<213> Medicago truncatula

<400> 96

Met Val Asn Ile Leu Lys Phe Ile Tyr Val Ile Ile Phe Phe Ile Leu

1 5 10 15

Met Phe Phe Val Leu Ile Asp Val Asp Gly His Val Leu Val Glu Cys

20 25 30

Ile Glu Asn Arg Asp Cys Glu Lys Gly Met Cys Lys Phe Pro Phe Ile

35 40 45

Val Arg Cys Leu Met Asp Gln Cys Lys Cys Val Arg Ile His Asn Leu

50 55 60

Ile

65

<210> 97

<211> 74

<212> PROTEIN

<213> Medicago truncatula

<400> 97

Met Ile Ile Gln Phe Ser Ile Tyr Tyr Met Gln Arg Arg Lys Leu Asn

1 5 10 15

Met Val Glu Ile Leu Lys Phe Ser His Ala Leu Ile Ile Phe Leu Phe

20 25 30

Leu Ser Ala Leu Val Thr Asn Ala Asn Ile Phe Phe Cys Ser Thr Asp

35 40 45

Glu Asp Cys Thr Trp Asn Leu Cys Arg Gln Pro Trp Val Gln Lys Cys

50 55 60

Arg Leu His Met Cys Ser Cys Glu Lys Asn

65 70

<210> 98

<211> 58

<212> PROTEIN

<213> Medicago truncatula

<400> 98

Met Asp Glu Val Phe Lys Phe Val Tyr Val Met Ile Ile Phe Pro Phe

1 5 10 15

Leu Ile Leu Asp Val Ala Thr Asn Ala Glu Lys Ile Arg Arg Cys Phe

20 25 30

Asn Asp Ala His Cys Pro Pro Asp Met Cys Thr Leu Gly Val Ile Pro

35 40 45

Lys Cys Ser Arg Phe Thr Ile Cys Ile Cys

50 55

<210> 99

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 99

Met His Arg Lys Pro Asn Met Thr Lys Phe Phe Lys Phe Val Tyr Thr

1 5 10 15

Met Phe Ile Leu Ile Ser Leu Phe Leu Val Val Thr Asn Ala Asn Ala

20 25 30

Asn Asn Cys Thr Asp Thr Ser Asp Cys Ser Ser Asn His Cys Ser Tyr

35 40 45

Glu Gly Val Ser Leu Cys Met Asn Gly Gln Cys Ile Cys Ile Tyr Glu

50 55 60

<210> 100

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 100

Met Gln Met Lys Lys Met Ala Thr Ile Leu Lys Phe Val Tyr Leu Ile

1 5 10 15

Ile Leu Leu Ile Tyr Pro Leu Leu Val Val Thr Glu Glu Ser His Tyr

20 25 30

Met Lys Phe Ser Ile Cys Lys Asp Asp Thr Asp Cys Pro Thr Leu Phe

35 40 45

Cys Val Leu Pro Asn Val Pro Lys Cys Ile Gly Ser Lys Cys His Cys

50 55 60

Lys Leu Met Val Asn

65

<210> 101

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 101

Met Val Glu Thr Leu Arg Leu Phe Tyr Ile Met Ile Leu Phe Val Ser

1 5 10 15

Leu Tyr Leu Val Val Val Asp Gly Val Ser Lys Leu Ala Gln Ser Cys

20 25 30

Ser Glu Asp Phe Glu Cys Tyr Ile Lys Asn Pro His Ala Pro Phe Gly

35 40 45

Gln Leu Arg Cys Phe Glu Gly Tyr Cys Gln Arg Leu Asp Lys Pro Thr

50 55 60

<210> 102

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 102

Met Thr Thr Phe Leu Lys Val Ala Tyr Ile Met Ile Ile Cys Val Phe

1 5 10 15

Val Leu His Leu Ala Ala Gln Val Asp Ser Gln Lys Arg Leu His Gly

20 25 30

Cys Lys Glu Asp Arg Asp Cys Asp Asn Ile Cys Ser Val His Ala Val

35 40 45

Thr Lys Cys Ile Gly Asn Met Cys Arg Cys Leu Ala Asn Val Lys

50 55 60

<210> 103

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 103

Met Arg Ile Asn Arg Thr Pro Ala Ile Phe Lys Phe Val Tyr Thr Ile

1 5 10 15

Ile Ile Tyr Leu Phe Leu Leu Arg Val Val Ala Lys Asp Leu Pro Phe

20 25 30

Asn Ile Cys Glu Lys Asp Glu Asp Cys Leu Glu Phe Cys Ala His Asp

35 40 45

Lys Val Ala Lys Cys Met Leu Asn Ile Cys Phe Cys Phe

50 55 60

<210> 104

<211> 54

<212> PROTEIN

<213> Medicago truncatula

<400> 104

Met Ala Glu Ile Leu Lys Ile Leu Tyr Val Phe Ile Ile Phe Leu Ser

1 5 10 15

Leu Ile Leu Ala Val Ile Ser Gln His Pro Phe Thr Pro Cys Glu Thr

20 25 30

Asn Ala Asp Cys Lys Cys Arg Asn His Lys Arg Pro Asp Cys Leu Trp

35 40 45

His Lys Cys Tyr Cys Tyr

50

<210> 105

<211> 65

<212> PROTEIN

<213> Medicago truncatula

<400> 105

Met Arg Lys Ser Met Ala Thr Ile Leu Lys Phe Val Tyr Val Ile Met

1 5 10 15

Leu Phe Ile Tyr Ser Leu Phe Val Ile Glu Ser Phe Gly His Arg Phe

20 25 30

Leu Ile Tyr Asn Asn Cys Lys Asn Asp Thr Glu Cys Pro Asn Asp Cys

35 40 45

Gly Pro His Glu Gln Ala Lys Cys Ile Leu Tyr Ala Cys Tyr Cys Val

50 55 60

Glu

65

<210> 106

<211> 58

<212> PROTEIN

<213> Medicago truncatula

<400> 106

Met Asn Thr Ile Leu Lys Phe Ile Phe Val Val Phe Leu Phe Leu Ser

1 5 10 15

Ile Phe Leu Ser Ala Gly Asn Ser Lys Ser Tyr Gly Pro Cys Thr Thr

20 25 30

Leu Gln Asp Cys Glu Thr His Asn Trp Phe Glu Val Cys Ser Cys Ile

35 40 45

Asp Phe Glu Cys Lys Cys Trp Ser Leu Leu

50 55

<210> 107

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 107

Met Ala Glu Ile Ile Lys Phe Val Tyr Ile Met Ile Leu Cys Val Ser

1 5 10 15

Leu Leu Leu Ile Ala Glu Ala Ser Gly Lys Glu Cys Val Thr Asp Ala

20 25 30

Asp Cys Glu Asn Leu Tyr Pro Gly Asn Lys Lys Pro Met Phe Cys Asn

35 40 45

Asn Thr Gly Tyr Cys Met Ser Leu Tyr Lys Glu Pro Ser Arg Tyr Met

50 55 60

<210> 108

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 108

Met Ala Lys Ile Ile Lys Phe Val Tyr Ile Met Ile Leu Cys Val Ser

1 5 10 15

Leu Leu Leu Ile Val Glu Ala Gly Gly Lys Glu Cys Val Thr Asp Val

20 25 30

Asp Cys Glu Lys Ile Tyr Pro Gly Asn Lys Lys Pro Leu Ile Cys Ser

35 40 45

Thr Gly Tyr Cys Tyr Ser Leu Tyr Glu Glu Pro Pro Arg Tyr His Lys

50 55 60

<210> 109

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 109

Met Ala Lys Val Thr Lys Phe Gly Tyr Ile Ile Ile His Phe Leu Ser

1 5 10 15

Leu Phe Phe Leu Ala Met Asn Val Ala Gly Gly Arg Glu Cys His Ala

20 25 30

Asn Ser His Cys Val Gly Lys Ile Thr Cys Val Leu Pro Gln Lys Pro

35 40 45

Glu Cys Trp Asn Tyr Ala Cys Val Cys Tyr Asp Ser Asn Lys Tyr Arg

50 55 60

<210> 110

<211> 55

<212> PROTEIN

<213> Medicago truncatula

<400> 110

Met Ala Lys Ile Phe Asn Tyr Val Tyr Ala Leu Ile Met Phe Leu Ser

1 5 10 15

Leu Phe Leu Met Gly Thr Ser Gly Met Lys Asn Gly Cys Lys His Thr

20 25 30

Gly His Cys Pro Arg Lys Met Cys Gly Ala Lys Thr Thr Lys Cys Arg

35 40 45

Asn Asn Lys Cys Gln Cys Val

50 55

<210> 111

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 111

Met Thr Glu Ile Leu Lys Phe Val Cys Val Met Ile Ile Phe Ile Ser

1 5 10 15

Ser Phe Ile Val Ser Lys Ser Leu Asn Gly Gly Gly Lys Asp Lys Cys

20 25 30

Phe Arg Asp Ser Asp Cys Pro Lys His Met Cys Pro Ser Ser Leu Val

35 40 45

Ala Lys Cys Ile Asn Arg Leu Cys Arg Cys Arg Arg Pro Glu Leu Gln

50 55 60

Val Gln Leu Asn Pro

65

<210> 112

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 112

Met Ala His Ile Ile Met Phe Val Tyr Ala Leu Ile Tyr Ala Leu Ile

1 5 10 15

Ile Phe Ser Ser Leu Phe Val Arg Asp Gly Ile Pro Cys Leu Ser Asp

20 25 30

Asp Glu Cys Pro Glu Met Ser His Tyr Ser Phe Lys Cys Asn Asn Lys

35 40 45

Ile Cys Glu Tyr Asp Leu Gly Glu Met Ser Asp Asp Asp Tyr Tyr Leu

50 55 60

Glu Met Ser Arg Glu

65

<210> 113

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 113

Met Tyr Arg Glu Lys Asn Met Ala Lys Thr Leu Lys Phe Val Tyr Val

1 5 10 15

Ile Val Leu Phe Leu Ser Leu Phe Leu Ala Ala Lys Asn Ile Asp Gly

20 25 30

Arg Val Ser Tyr Asn Ser Phe Ile Ala Leu Pro Val Cys Gln Thr Ala

35 40 45

Ala Asp Cys Pro Glu Gly Thr Arg Gly Arg Thr Tyr Lys Cys Ile Asn

50 55 60

Asn Lys Cys Arg Tyr Pro Lys Leu Leu Lys Pro Ile Gln

65 70 75

<210> 114

<211> 56

<212> PROTEIN

<213> Medicago truncatula

<400> 114

Met Ala His Ile Phe Asn Tyr Val Tyr Ala Leu Leu Val Phe Leu Ser

1 5 10 15

Leu Phe Leu Met Val Thr Asn Gly Ile His Ile Gly Cys Asp Lys Asp

20 25 30

Arg Asp Cys Pro Lys Gln Met Cys His Leu Asn Gln Thr Pro Lys Cys

35 40 45

Leu Lys Asn Ile Cys Lys Cys Val

50 55

<210> 115

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 115

Met Ala Glu Ile Leu Lys Cys Phe Tyr Thr Met Asn Leu Phe Ile Phe

1 5 10 15

Leu Ile Ile Leu Pro Ala Lys Ile Arg Glu His Ile Gln Cys Val Ile

20 25 30

Asp Asp Asp Cys Pro Lys Ser Leu Asn Lys Leu Leu Ile Ile Lys Cys

35 40 45

Ile Asn His Val Cys Gln Tyr Val Gly Asn Leu Pro Asp Phe Ala Ser

50 55 60

Gln Ile Pro Lys Ser Thr Lys Met Pro Tyr Lys Gly Glu

65 70 75

<210> 116

<211> 70

<212> PROTEIN

<213> Medicago truncatula

<400> 116

Met Ala Tyr Ile Ser Arg Ile Phe Tyr Val Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe Phe Val Val Ile Asn Gly Val Lys Ser Leu Leu Leu Ile Lys

20 25 30

Val Arg Ser Phe Ile Pro Cys Gln Arg Ser Asp Asp Cys Pro Arg Asn

35 40 45

Leu Cys Val Asp Gln Ile Ile Pro Thr Cys Val Trp Ala Lys Cys Lys

50 55 60

Cys Lys Asn Tyr Asn Asp

65 70

<210> 117

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 117

Met Ala Asn Val Thr Lys Phe Val Tyr Ile Ala Ile Tyr Phe Leu Ser

1 5 10 15

Leu Phe Phe Ile Ala Lys Asn Asp Ala Thr Ala Thr Phe Cys His Asp

20 25 30

Asp Ser His Cys Val Thr Lys Ile Lys Cys Val Leu Pro Arg Thr Pro

35 40 45

Gln Cys Arg Asn Glu Ala Cys Gly Cys Tyr His Ser Asn Lys Phe Arg

50 55 60

<210> 118

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 118

Met Gly Glu Ile Met Lys Phe Val Tyr Val Met Ile Ile Tyr Leu Phe

1 5 10 15

Met Phe Asn Val Ala Thr Gly Ser Glu Phe Ile Phe Thr Lys Lys Leu

20 25 30

Thr Ser Cys Asp Ser Ser Lys Asp Cys Arg Ser Phe Leu Cys Tyr Ser

35 40 45

Pro Lys Phe Pro Val Cys Lys Arg Gly Ile Cys Glu Cys Ile

50 55 60

<210> 119

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 119

Met Gly Glu Met Phe Lys Phe Ile Tyr Thr Phe Ile Leu Phe Val His

1 5 10 15

Leu Phe Leu Val Val Ile Phe Glu Asp Ile Gly His Ile Lys Tyr Cys

20 25 30

Gly Ile Val Asp Asp Cys Tyr Lys Ser Lys Lys Pro Leu Phe Lys Ile

35 40 45

Trp Lys Cys Val Glu Asn Val Cys Val Leu Trp Tyr Lys

50 55 60

<210> 120

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 120

Met Ala Arg Thr Leu Lys Phe Val Tyr Ser Met Ile Leu Phe Leu Ser

1 5 10 15

Leu Phe Leu Val Ala Asn Gly Leu Lys Ile Phe Cys Ile Asp Val Ala

20 25 30

Asp Cys Pro Lys Asp Leu Tyr Pro Leu Leu Tyr Lys Cys Ile Tyr Asn

35 40 45

Lys Cys Ile Val Phe Thr Arg Ile Pro Phe Pro Phe Asp Trp Ile

50 55 60

<210> 121

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 121

Met Ala Asn Ile Thr Lys Phe Val Tyr Ile Ala Ile Leu Phe Leu Ser

1 5 10 15

Leu Phe Phe Ile Gly Met Asn Asp Ala Ala Ile Leu Glu Cys Arg Glu

20 25 30

Asp Ser His Cys Val Thr Lys Ile Lys Cys Val Leu Pro Arg Lys Pro

35 40 45

Glu Cys Arg Asn Asn Ala Cys Thr Cys Tyr Lys Gly Gly Phe Ser Phe

50 55 60

His His

65

<210> 122

<211> 68

<212> PROTEIN

<213> Medicago truncatula

<400> 122

Met Gln Arg Val Lys Lys Met Ser Glu Thr Leu Lys Phe Val Tyr Val

1 5 10 15

Leu Ile Leu Phe Ile Ser Ile Phe His Val Val Ile Val Cys Asp Ser

20 25 30

Ile Tyr Phe Pro Val Ser Arg Pro Cys Ile Thr Asp Lys Asp Cys Pro

35 40 45

Asn Met Lys His Tyr Lys Ala Lys Cys Arg Lys Gly Phe Cys Ile Ser

50 55 60

Ser Arg Val Arg

65

<210> 123

<211> 72

<212> PROTEIN

<213> Medicago truncatula

<400> 123

Met Gln Ile Arg Lys Ile Met Ser Gly Val Leu Lys Phe Val Tyr Ala

1 5 10 15

Ile Ile Leu Phe Leu Phe Leu Phe Leu Val Ala Arg Glu Val Gly Gly

20 25 30

Leu Glu Thr Ile Glu Cys Glu Thr Asp Gly Asp Cys Pro Arg Ser Met

35 40 45

Ile Lys Met Trp Asn Lys Asn Tyr Arg His Lys Cys Ile Asp Gly Lys

50 55 60

Cys Glu Trp Ile Lys Lys Leu Pro

65 70

<210> 124

<211> 54

<212> PROTEIN

<213> Medicago truncatula

<400> 124

Met Phe Val Tyr Asp Leu Ile Leu Phe Ile Ser Leu Ile Leu Val Val

1 5 10 15

Thr Gly Ile Asn Ala Glu Ala Asp Thr Ser Cys His Ser Phe Asp Asp

20 25 30

Cys Pro Trp Val Ala His His Tyr Arg Glu Cys Ile Glu Gly Leu Cys

35 40 45

Ala Tyr Arg Ile Leu Tyr

50

<210> 125

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 125

Met Gln Arg Arg Lys Lys Ser Met Ala Lys Met Leu Lys Phe Phe Phe

1 5 10 15

Ala Ile Ile Leu Leu Leu Ser Leu Phe Leu Val Ala Thr Glu Val Gly

20 25 30

Gly Ala Tyr Ile Glu Cys Glu Val Asp Asp Asp Cys Pro Lys Pro Met

35 40 45

Lys Asn Ser His Pro Asp Thr Tyr Tyr Lys Cys Val Lys His Arg Cys

50 55 60

Gln Trp Ala Trp Lys

65

<210> 126

<211> 140

<212> PROTEIN

<213> Medicago truncatula

<400> 126

Met Phe Val Tyr Thr Leu Ile Ile Phe Leu Phe Pro Ser His Val Ile

1 5 10 15

Thr Asn Lys Ile Ala Ile Tyr Cys Val Ser Asp Asp Asp Cys Leu Lys

20 25 30

Thr Phe Thr Pro Leu Asp Leu Lys Cys Val Asp Asn Val Cys Glu Phe

35 40 45

Asn Leu Arg Cys Lys Gly Lys Cys Gly Glu Arg Asp Glu Lys Phe Val

50 55 60

Phe Leu Lys Ala Leu Lys Lys Met Asp Gln Lys Leu Val Leu Glu Glu

65 70 75 80

Gln Gly Asn Ala Arg Glu Val Lys Ile Pro Lys Lys Leu Leu Phe Asp

85 90 95

Arg Ile Gln Val Pro Thr Pro Ala Thr Lys Asp Gln Val Glu Glu Asp

100 105 110

Asp Tyr Asp Asp Asp Asp Glu Glu Glu Glu Glu Glu Glu Asp Asp Val

115 120 125

Asp Met Trp Phe His Leu Pro Asp Val Val Cys His

130 135 140

<210> 127

<211> 60

<212> PROTEIN

<213> Medicago truncatula

<400> 127

Met Ala Lys Phe Ser Met Phe Val Tyr Ala Leu Ile Asn Phe Leu Ser

1 5 10 15

Leu Phe Leu Val Glu Thr Ala Ile Thr Asn Ile Arg Cys Val Ser Asp

20 25 30

Asp Asp Cys Pro Lys Val Ile Lys Pro Leu Val Met Lys Cys Ile Gly

35 40 45

Asn Tyr Cys Tyr Phe Phe Met Ile Tyr Glu Gly Pro

50 55 60

<210> 128

<211> 58

<212> PROTEIN

<213> Medicago truncatula

<400> 128

Met Ala His Lys Phe Val Tyr Ala Ile Ile Leu Phe Ile Phe Leu Phe

1 5 10 15

Leu Val Ala Lys Asn Val Lys Gly Tyr Val Val Cys Arg Thr Val Asp

20 25 30

Asp Cys Pro Pro Asp Thr Arg Asp Leu Arg Tyr Arg Cys Leu Asn Gly

35 40 45

Lys Cys Lys Ser Tyr Arg Leu Ser Tyr Gly

50 55

<210> 129

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 129

Met Gln Arg Lys Lys Asn Met Gly Gln Ile Leu Ile Phe Val Phe Ala

1 5 10 15

Leu Ile Asn Phe Leu Ser Pro Ile Leu Val Glu Met Thr Thr Thr Thr

20 25 30

Ile Pro Cys Thr Phe Ile Asp Asp Cys Pro Lys Met Pro Leu Val Val

35 40 45

Lys Cys Ile Asp Asn Phe Cys Asn Tyr Phe Glu Ile Lys

50 55 60

<210> 130

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 130

Met Ala Gln Thr Leu Met Leu Val Tyr Ala Leu Ile Ile Phe Thr Ser

1 5 10 15

Leu Phe Leu Val Val Ile Ser Arg Gln Thr Asp Ile Pro Cys Lys Ser

20 25 30

Asp Asp Ala Cys Pro Arg Val Ser Ser His His Ile Glu Cys Val Lys

35 40 45

Gly Phe Cys Thr Tyr Trp Lys Leu Asp

50 55

<210> 131

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 131

Met Leu Arg Arg Lys Asn Thr Val Gln Ile Leu Met Phe Val Ser Ala

1 5 10 15

Leu Leu Ile Tyr Ile Phe Leu Phe Leu Val Ile Thr Ser Ser Ala Asn

20 25 30

Ile Pro Cys Asn Ser Asp Ser Asp Cys Pro Trp Lys Ile Tyr Tyr Thr

35 40 45

Tyr Arg Cys Asn Asp Gly Phe Cys Val Tyr Lys Ser Ile Asp Pro Ser

50 55 60

Thr Ile Pro Gln Tyr Met Thr Asp Leu Ile Phe Pro Arg

65 70 75

<210> 132

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 132

Met Ala Val Ile Leu Lys Phe Val Tyr Ile Met Ile Ile Phe Leu Phe

1 5 10 15

Leu Leu Tyr Val Val Asn Gly Thr Arg Cys Asn Arg Asp Glu Asp Cys

20 25 30

Pro Phe Ile Cys Thr Gly Pro Gln Ile Pro Lys Cys Val Ser His Ile

35 40 45

Cys Phe Cys Leu Ser Ser Gly Lys Glu Ala Tyr

50 55

<210> 133

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 133

Met Asp Ala Ile Leu Lys Phe Ile Tyr Ala Met Phe Leu Phe Leu Phe

1 5 10 15

Leu Phe Val Thr Thr Arg Asn Val Glu Ala Leu Phe Glu Cys Asn Arg

20 25 30

Asp Phe Val Cys Gly Asn Asp Asp Glu Cys Val Tyr Pro Tyr Ala Val

35 40 45

Gln Cys Ile His Arg Tyr Cys Lys Cys Leu Lys Ser Arg Asn

50 55 60

<210> 134

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 134

Met Gln Ile Gly Arg Lys Lys Met Gly Glu Thr Pro Lys Leu Val Tyr

1 5 10 15

Val Ile Ile Leu Phe Leu Ser Ile Phe Leu Cys Thr Asn Ser Ser Phe

20 25 30

Ser Gln Met Ile Asn Phe Arg Gly Cys Lys Arg Asp Lys Asp Cys Pro

35 40 45

Gln Phe Arg Gly Val Asn Ile Arg Cys Arg Ser Gly Phe Cys Thr Pro

50 55 60

Ile Asp Ser

65

<210> 135

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 135

Met Gln Met Arg Lys Asn Met Ala Gln Ile Leu Phe Tyr Val Tyr Ala

1 5 10 15

Leu Leu Ile Leu Phe Ser Pro Phe Leu Val Ala Arg Ile Met Val Val

20 25 30

Asn Pro Asn Asn Pro Cys Val Thr Asp Ala Asp Cys Gln Arg Tyr Arg

35 40 45

His Lys Leu Ala Thr Arg Met Val Cys Asn Ile Gly Phe Cys Leu Met

50 55 60

Asp Phe Thr His Asp Pro Tyr Ala Pro Ser Leu Pro

65 70 75

<210> 136

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 136

Met Tyr Val Tyr Tyr Ile Gln Met Gly Lys Asn Met Ala Gln Arg Phe

1 5 10 15

Met Phe Ile Tyr Ala Leu Ile Ile Phe Leu Ser Gln Phe Phe Val Val

20 25 30

Ile Asn Thr Ser Asp Ile Pro Asn Asn Ser Asn Arg Asn Ser Pro Lys

35 40 45

Glu Asp Val Phe Cys Asn Ser Asn Asp Asp Cys Pro Thr Ile Leu Tyr

50 55 60

Tyr Val Ser Lys Cys Val Tyr Asn Phe Cys Glu Tyr Trp

65 70 75

<210> 137

<211> 67

<212> PROTEIN

<213> Medicago truncatula

<400> 137

Met Ala Lys Ile Val Asn Phe Val Tyr Ser Met Ile Ile Phe Val Ser

1 5 10 15

Leu Phe Leu Val Ala Thr Lys Gly Gly Ser Lys Pro Phe Leu Thr Arg

20 25 30

Pro Tyr Pro Cys Asn Thr Gly Ser Asp Cys Pro Gln Asn Met Cys Pro

35 40 45

Pro Gly Tyr Lys Pro Gly Cys Glu Asp Gly Tyr Cys Asn His Cys Tyr

50 55 60

Lys Arg Trp

65

<210> 138

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 138

Met Val Arg Thr Leu Lys Phe Val Tyr Val Ile Ile Leu Ile Leu Ser

1 5 10 15

Leu Phe Leu Val Ala Lys Gly Gly Gly Lys Lys Ile Tyr Cys Glu Asn

20 25 30

Ala Ala Ser Cys Pro Arg Leu Met Tyr Pro Leu Val Tyr Lys Cys Leu

35 40 45

Asp Asn Lys Cys Val Lys Phe Met Met Lys Ser Arg Phe Val

50 55 60

<210> 139

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 139

Met Ala Arg Thr Leu Lys Phe Val Tyr Ala Val Ile Leu Phe Leu Ser

1 5 10 15

Leu Phe Leu Val Ala Lys Gly Asp Asp Val Lys Ile Lys Cys Val Val

20 25 30

Ala Ala Asn Cys Pro Asp Leu Met Tyr Pro Leu Val Tyr Lys Cys Leu

35 40 45

Asn Gly Ile Cys Val Gln Phe Thr Leu Thr Phe Pro Phe Val

50 55 60

<210> 140

<211> 65

<212> PROTEIN

<213> Medicago truncatula

<400> 140

Met Ser Asn Thr Leu Met Phe Val Ile Thr Phe Ile Val Leu Val Thr

1 5 10 15

Leu Phe Leu Gly Pro Lys Asn Val Tyr Ala Phe Gln Pro Cys Val Thr

20 25 30

Thr Ala Asp Cys Met Lys Thr Leu Lys Thr Asp Glu Asn Ile Trp Tyr

35 40 45

Glu Cys Ile Asn Asp Phe Cys Ile Pro Phe Pro Ile Pro Lys Gly Arg

50 55 60

Lys

65

<210> 141

<211> 76

<212> PROTEIN

<213> Medicago truncatula

<400> 141

Met Lys Arg Val Val Asn Met Ala Lys Ile Val Lys Tyr Val Tyr Val

1 5 10 15

Ile Ile Ile Phe Leu Ser Leu Phe Leu Val Ala Thr Lys Ile Glu Gly

20 25 30

Tyr Tyr Tyr Lys Cys Phe Lys Asp Ser Asp Cys Val Lys Leu Leu Cys

35 40 45

Arg Ile Pro Leu Arg Pro Lys Cys Met Tyr Arg His Ile Cys Lys Cys

50 55 60

Lys Val Val Leu Thr Gln Asn Asn Tyr Val Leu Thr

65 70 75

<210> 142

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 142

Met Lys Arg Gly Lys Asn Met Ser Lys Ile Leu Lys Phe Ile Tyr Ala

1 5 10 15

Thr Leu Val Leu Tyr Leu Phe Leu Val Val Thr Lys Ala Ser Asp Asp

20 25 30

Glu Cys Lys Ile Asp Gly Asp Cys Pro Ile Ser Trp Gln Lys Phe His

35 40 45

Thr Tyr Lys Cys Ile Asn Gln Lys Cys Lys Trp Val Leu Arg Phe His

50 55 60

Glu Tyr

65

<210> 143

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 143

Met Ala Lys Thr Leu Asn Phe Met Phe Ala Leu Ile Leu Phe Ile Ser

1 5 10 15

Leu Phe Leu Val Ser Lys Asn Val Ala Ile Asp Ile Phe Val Cys Gln

20 25 30

Thr Asp Ala Asp Cys Pro Lys Ser Glu Leu Ser Met Tyr Thr Trp Lys

35 40 45

Cys Ile Asp Asn Glu Cys Asn Leu Phe Lys Val Met Gln Gln Met Val

50 55 60

<210> 144

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 144

Met Ala Asn Thr His Lys Leu Val Ser Met Ile Leu Phe Ile Phe Leu

1 5 10 15

Phe Leu Val Ala Asn Asn Val Glu Gly Tyr Val Asn Cys Glu Thr Asp

20 25 30

Ala Asp Cys Pro Pro Ser Thr Arg Val Lys Arg Phe Lys Cys Val Lys

35 40 45

Gly Glu Cys Arg Trp Thr Arg Met Ser Tyr Ala

50 55

<210> 145

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 145

Met Ala His Phe Leu Met Phe Val Tyr Ala Leu Ile Thr Cys Leu Ser

1 5 10 15

Leu Phe Leu Val Glu Met Gly His Leu Ser Ile His Cys Val Ser Val

20 25 30

Asp Asp Cys Pro Lys Val Glu Lys Pro Ile Thr Met Lys Cys Ile Asn

35 40 45

Asn Tyr Cys Lys Tyr Phe Val Asp His Lys Leu

50 55

<210> 146

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 146

Met Asn Gln Ile Pro Met Phe Gly Tyr Thr Leu Ile Ile Phe Phe Ser

1 5 10 15

Leu Phe Pro Val Ile Thr Asn Gly Asp Arg Ile Pro Cys Val Thr Asn

20 25 30

Gly Asp Cys Pro Val Met Arg Leu Pro Leu Tyr Met Arg Cys Ile Thr

35 40 45

Tyr Ser Cys Glu Leu Phe Phe Asp Gly Pro Asn Leu Cys Ala Val Glu

50 55 60

Arg Ile

65

<210> 147

<211> 61

<212> PROTEIN

<213> Medicago truncatula

<400> 147

Met Arg Lys Asp Met Ala Arg Ile Ser Leu Phe Val Tyr Ala Leu Ile

1 5 10 15

Ile Phe Phe Ser Leu Phe Phe Val Leu Thr Asn Gly Glu Leu Glu Ile

20 25 30

Arg Cys Val Ser Asp Ala Asp Cys Pro Leu Phe Pro Leu Pro Leu His

35 40 45

Asn Arg Cys Ile Asp Asp Val Cys His Leu Phe Thr Ser

50 55 60

<210> 148

<211> 60

<212> PROTEIN

<213> Medicago truncatula

<400> 148

Met Ala Gln Ile Leu Met Phe Val Tyr Phe Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe Leu Val Glu Ser Ile Lys Ile Phe Thr Glu His Arg Cys Arg

20 25 30

Thr Asp Ala Asp Cys Pro Ala Arg Glu Leu Pro Glu Tyr Leu Lys Cys

35 40 45

Gln Gly Gly Met Cys Arg Leu Leu Ile Lys Lys Asp

50 55 60

<210> 149

<211> 56

<212> PROTEIN

<213> Medicago truncatula

<400> 149

Met Ala Arg Val Ile Ser Leu Phe Tyr Ala Leu Ile Ile Phe Leu Phe

1 5 10 15

Leu Phe Leu Val Ala Thr Asn Gly Asp Leu Ser Pro Cys Leu Arg Ser

20 25 30

Gly Asp Cys Ser Lys Asp Glu Cys Pro Ser His Leu Val Pro Lys Cys

35 40 45

Ile Gly Leu Thr Cys Tyr Cys Ile

50 55

<210> 150

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 150

Met Gln Arg Arg Lys Asn Met Ala Gln Ile Leu Leu Phe Ala Tyr Val

1 5 10 15

Phe Ile Ile Ser Ile Ser Leu Phe Leu Val Val Thr Asn Gly Val Lys

20 25 30

Ile Pro Cys Val Lys Asp Thr Asp Cys Pro Thr Leu Pro Cys Pro Leu

35 40 45

Tyr Ser Lys Cys Val Asp Gly Phe Cys Lys Met Leu Ser Ile

50 55 60

<210> 151

<211> 66

<212> PROTEIN

<213> Medicago truncatula

<400> 151

Met Asn His Ile Ser Lys Phe Val Tyr Ala Leu Ile Ile Phe Leu Ser

1 5 10 15

Val Tyr Leu Val Val Leu Asp Gly Arg Pro Val Ser Cys Lys Asp His

20 25 30

Tyr Asp Cys Arg Arg Lys Val Lys Ile Val Gly Cys Ile Phe Pro Gln

35 40 45

Glu Lys Pro Met Cys Ile Asn Ser Met Cys Thr Cys Ile Arg Glu Ile

50 55 60

Val Pro

65

<210> 152

<211> 86

<212> PROTEIN

<213> Medicago truncatula

<400> 152

Met Lys Ser Gln Asn His Ala Lys Phe Ile Ser Phe Tyr Lys Asn Asp

1 5 10 15

Leu Phe Lys Ile Phe Gln Asn Asn Asp Ser His Phe Lys Val Phe Phe

20 25 30

Ala Leu Ile Ile Phe Leu Tyr Thr Tyr Leu His Val Thr Asn Gly Val

35 40 45

Phe Val Ser Cys Asn Ser His Ile His Cys Arg Val Asn Asn His Lys

50 55 60

Ile Gly Cys Asn Ile Pro Glu Gln Tyr Leu Leu Cys Val Asn Leu Phe

65 70 75 80

Cys Leu Trp Leu Asp Tyr

85

<210> 153

<211> 62

<212> PROTEIN

<213> Medicago truncatula

<400> 153

Met Thr Tyr Ile Ser Lys Val Val Tyr Ala Leu Ile Ile Phe Leu Ser

1 5 10 15

Ile Tyr Val Gly Val Asn Asp Cys Met Leu Val Thr Cys Glu Asp His

20 25 30

Phe Asp Cys Arg Gln Asn Val Gln Gln Val Gly Cys Ser Phe Arg Glu

35 40 45

Ile Pro Gln Cys Ile Asn Ser Ile Cys Lys Cys Met Lys Gly

50 55 60

<210> 154

<211> 63

<212> PROTEIN

<213> Medicago truncatula

<400> 154

Met Thr His Ile Ser Lys Phe Val Phe Ala Leu Ile Ile Phe Leu Ser

1 5 10 15

Ile Tyr Val Gly Val Asn Asp Cys Lys Arg Ile Pro Cys Lys Asp Asn

20 25 30

Asn Asp Cys Asn Asn Asn Trp Gln Leu Leu Ala Cys Arg Phe Glu Arg

35 40 45

Glu Val Pro Arg Cys Ile Asn Ser Ile Cys Lys Cys Met Pro Met

50 55 60

<210> 155

<211> 60

<212> PROTEIN

<213> Medicago truncatula

<400> 155

Met Val Gln Thr Pro Lys Leu Val Tyr Val Ile Val Leu Leu Leu Ser

1 5 10 15

Ile Phe Leu Gly Met Thr Ile Cys Asn Ser Ser Phe Ser His Phe Phe

20 25 30

Glu Gly Ala Cys Lys Ser Asp Lys Asp Cys Pro Lys Leu His Arg Ser

35 40 45

Asn Val Arg Cys Arg Lys Gly Gln Cys Val Gln Ile

50 55 60

<210> 156

<211> 77

<212> PROTEIN

<213> Medicago truncatula

<400> 156

Met Thr Lys Ile Leu Met Leu Phe Tyr Ala Met Ile Val Phe His Ser

1 5 10 15

Ile Phe Leu Val Ala Ser Tyr Thr Asp Glu Cys Ser Thr Asp Ala Asp

20 25 30

Cys Glu Tyr Ile Leu Cys Leu Phe Pro Ile Ile Lys Arg Cys Ile His

35 40 45

Asn His Cys Lys Cys Val Pro Met Gly Ser Ile Glu Pro Met Ser Thr

50 55 60

Ile Pro Asn Gly Val His Lys Phe His Ile Ile Asn Asn

65 70 75

<210> 157

<211> 64

<212> PROTEIN

<213> Medicago truncatula

<400> 157

Met Ala Lys Thr Leu Asn Phe Val Cys Ala Met Ile Leu Phe Ile Ser

1 5 10 15

Leu Phe Leu Val Ser Lys Asn Val Ala Leu Tyr Ile Ile Glu Cys Lys

20 25 30

Thr Asp Ala Asp Cys Pro Ile Ser Lys Leu Asn Met Tyr Asn Trp Arg

35 40 45

Cys Ile Lys Ser Ser Cys His Leu Tyr Lys Val Ile Gln Phe Met Val

50 55 60

<210> 158

<211> 72

<212> PROTEIN

<213> Medicago truncatula

<400> 158

Met Gln Lys Glu Lys Asn Met Ala Lys Thr Phe Glu Phe Val Tyr Ala

1 5 10 15

Met Ile Ile Phe Ile Leu Leu Phe Leu Val Glu Asn Asn Phe Ala Ala

20 25 30

Tyr Ile Ile Glu Cys Gln Thr Asp Asp Asp Cys Pro Lys Ser Gln Leu

35 40 45

Glu Met Phe Ala Trp Lys Cys Val Lys Asn Gly Cys His Leu Phe Gly

50 55 60

Met Tyr Glu Asp Asp Asp Asp Pro

65 70

<210> 159

<211> 57

<212> PROTEIN

<213> Medicago truncatula

<400> 159

Met Ala Ala Thr Arg Lys Phe Ile Tyr Val Leu Ser His Phe Leu Phe

1 5 10 15

Leu Phe Leu Val Thr Lys Ile Thr Asp Ala Arg Val Cys Lys Ser Asp

20 25 30

Lys Asp Cys Lys Asp Ile Ile Ile Tyr Arg Tyr Ile Leu Lys Cys Arg

35 40 45

Asn Gly Glu Cys Val Lys Ile Lys Ile

50 55

<210> 160

<211> 75

<212> PROTEIN

<213> Medicago truncatula

<400> 160

Met Gln Arg Leu Asp Asn Met Ala Lys Asn Val Lys Phe Ile Tyr Val

1 5 10 15

Ile Ile Leu Leu Leu Phe Ile Phe Leu Val Ile Ile Val Cys Asp Ser

20 25 30

Ala Phe Val Pro Asn Ser Gly Pro Cys Thr Thr Asp Lys Asp Cys Lys

35 40 45

Gln Val Lys Gly Tyr Ile Ala Arg Cys Arg Lys Gly Tyr Cys Met Gln

50 55 60

Ser Val Lys Arg Thr Trp Ser Ser Tyr Ser Arg

65 70 75

<210> 161

<211> 102

<212> PROTEIN

<213> Medicago truncatula

<400> 161

Met Lys Phe Ile Tyr Ile Met Ile Leu Phe Leu Ser Leu Phe Leu Val

1 5 10 15

Gln Phe Leu Thr Cys Lys Gly Leu Thr Val Pro Cys Glu Asn Pro Thr

20 25 30

Thr Cys Pro Glu Asp Phe Cys Thr Pro Pro Met Ile Thr Arg Cys Ile

35 40 45

Asn Phe Ile Cys Leu Cys Asp Gly Pro Glu Tyr Ala Glu Pro Glu Tyr

50 55 60

Asp Gly Pro Glu Pro Glu Tyr Asp His Lys Gly Asp Phe Leu Ser Val

65 70 75 80

Lys Pro Lys Ile Ile Asn Glu Asn Met Met Met Arg Glu Arg His Met

85 90 95

Met Lys Glu Ile Glu Val

100

<210> 162

<211> 59

<212> PROTEIN

<213> Medicago truncatula

<400> 162

Met Ala Gln Phe Leu Met Phe Ile Tyr Val Leu Ile Ile Phe Leu Tyr

1 5 10 15

Leu Phe Tyr Val Glu Ala Ala Met Phe Glu Leu Thr Lys Ser Thr Ile

20 25 30

Arg Cys Val Thr Asp Ala Asp Cys Pro Asn Val Val Lys Pro Leu Lys

35 40 45

Pro Lys Cys Val Asp Gly Phe Cys Glu Tyr Thr

50 55

<210> 163

<211> 70

<212> PROTEIN

<213> Medicago truncatula

<400> 163

Met Lys Met Arg Ile His Met Ala Gln Ile Ile Met Phe Phe Tyr Ala

1 5 10 15

Leu Ile Ile Phe Leu Ser Pro Phe Leu Val Asp Arg Arg Ser Phe Pro

20 25 30

Ser Ser Phe Val Ser Pro Lys Ser Tyr Thr Ser Glu Ile Pro Cys Lys

35 40 45

Ala Thr Arg Asp Cys Pro Tyr Glu Leu Tyr Tyr Glu Thr Lys Cys Val

50 55 60

Asp Ser Leu Cys Thr Tyr

65 70

<210> 164

<211> 41

<212> PROTEIN

<213> Medicago truncatula

<400> 164

Thr Arg Met Leu Thr Ile Pro Cys Thr Ser Asp Asp Asn Cys Pro Lys

1 5 10 15

Val Ile Ser Pro Cys His Thr Lys Cys Phe Asp Gly Phe Cys Gly Trp

20 25 30

Tyr Ile Glu Gly Ser Tyr Glu Gly Pro

35 40

<210> 165

<211> 69

<212> PROTEIN

<213> Medicago truncatula

<400> 165

Met Ala Gln Phe Leu Leu Phe Val Tyr Ser Leu Ile Ile Phe Leu Ser

1 5 10 15

Leu Phe Phe Gly Glu Ala Ala Phe Glu Arg Thr Glu Thr Arg Met Leu

20 25 30

Thr Ile Pro Cys Thr Ser Asp Asp Asn Cys Pro Lys Val Ile Ser Pro

35 40 45

Cys His Thr Lys Cys Phe Asp Gly Phe Cys Gly Trp Tyr Ile Glu Gly

50 55 60

Ser Tyr Glu Gly Pro

65

<210> 166

<211> 78

<212> PROTEIN

<213> Buchnera aphidicola

<400> 166

Met Lys Leu Leu His Gly Phe Leu Ile Ile Met Leu Thr Met His Leu

1 5 10 15

Ser Ile Gln Tyr Ala Tyr Gly Gly Pro Phe Leu Thr Lys Tyr Leu Cys

20 25 30

Asp Arg Val Cys His Lys Leu Cys Gly Asp Glu Phe Val Cys Ser Cys

35 40 45

Ile Gln Tyr Lys Ser Leu Lys Gly Leu Trp Phe Pro His Cys Pro Thr

50 55 60

Gly Lys Ala Ser Val Val Leu His Asn Phe Leu Thr Ser Pro

65 70 75

<210> 167

<211> 77

<212> PROTEIN

<213> Buchnera aphidicola

<400> 167

Met Lys Leu Leu Tyr Gly Phe Leu Ile Ile Met Leu Thr Ile His Leu

1 5 10 15

Ser Val Gln Tyr Phe Glu Ser Pro Phe Glu Thr Lys Tyr Asn Cys Asp

20 25 30

Thr His Cys Asn Lys Leu Cys Gly Lys Ile Asp His Cys Ser Cys Ile

35 40 45

Gln Tyr His Ser Met Glu Gly Leu Trp Phe Pro His Cys Arg Thr Gly

50 55 60

Ser Ala Ala Gln Met Leu His Asp Phe Leu Ser Asn Pro

65 70 75

<210> 168

<211> 86

<212> PROTEIN

<213> Buchnera aphidicola

<400> 168

Met Ser Val Arg Lys Asn Val Leu Pro Thr Met Phe Val Val Leu Leu

1 5 10 15

Ile Met Ser Pro Val Thr Pro Thr Ser Val Phe Ile Ser Ala Val Cys

20 25 30

Tyr Ser Gly Cys Gly Ser Leu Ala Leu Val Cys Phe Val Ser Asn Gly

35 40 45

Ile Thr Asn Gly Leu Asp Tyr Phe Lys Ser Ser Ala Pro Leu Ser Thr

50 55 60

Ser Glu Thr Ser Cys Gly Glu Ala Phe Asp Thr Cys Thr Asp His Cys

65 70 75 80

Leu Ala Asn Phe Lys Phe

85

<210> 169

<211> 69

<212> PROTEIN

<213> Buchnera aphidicola

<400> 169

Met Arg Leu Leu Tyr Gly Phe Leu Ile Ile Met Leu Thr Ile Tyr Leu

1 5 10 15

Ser Val Gln Asp Phe Asp Pro Thr Glu Phe Lys Gly Pro Phe Pro Thr

20 25 30

Ile Glu Ile Cys Ser Lys Tyr Cys Ala Val Val Cys Asn Tyr Thr Ser

35 40 45

Arg Pro Cys Tyr Cys Val Glu Ala Ala Lys Glu Arg Asp Gln Trp Phe

50 55 60

Pro Tyr Cys Tyr Asp

65

<210> 170

<211> 77

<212> PROTEIN

<213> Buchnera aphidicola

<400> 170

Met Arg Leu Leu Tyr Gly Phe Leu Ile Ile Met Leu Thr Ile His Leu

1 5 10 15

Ser Val Gln Asp Ile Asp Pro Asn Thr Leu Arg Gly Pro Tyr Pro Thr

20 25 30

Lys Glu Ile Cys Ser Lys Tyr Cys Glu Tyr Asn Val Val Cys Gly Ala

35 40 45

Ser Leu Pro Cys Ile Cys Val Gln Asp Ala Arg Gln Leu Asp His Trp

50 55 60

Phe Ala Cys Cys Tyr Asp Gly Gly Pro Glu Met Leu Met

65 70 75

<210> 171

<211> 108

<212> PROTEIN

<213> Buchnera aphidicola

<400> 171

Met Lys Leu Phe Val Val Val Val Leu Val Ala Val Gly Ile Met Phe

1 5 10 15

Val Phe Ala Ser Asp Thr Ala Ala Ala Pro Thr Asp Tyr Glu Asp Thr

20 25 30

Asn Asp Met Ile Ser Leu Ser Ser Leu Val Gly Asp Asn Ser Pro Tyr

35 40 45

Val Arg Val Ser Ser Ala Asp Ser Gly Gly Ser Ser Lys Thr Ser Ser

50 55 60

Lys Asn Pro Ile Leu Gly Leu Leu Lys Ser Val Ile Lys Leu Leu Thr

65 70 75 80

Lys Ile Phe Gly Thr Tyr Ser Asp Ala Ala Pro Ala Met Pro Pro Ile

85 90 95

Pro Pro Ala Leu Arg Lys Asn Arg Gly Met Leu Ala

100 105

<210> 172

<211> 178

<212> PROTEIN

<213> Buchnera aphidicola

<400> 172

Met Val Ala Cys Lys Val Ile Leu Ala Val Ala Val Val Phe Val Ala

1 5 10 15

Ala Val Gln Gly Arg Pro Gly Gly Glu Pro Glu Trp Ala Ala Pro Ile

20 25 30

Phe Ala Glu Leu Lys Ser Val Ser Asp Asn Ile Thr Asn Leu Val Gly

35 40 45

Leu Asp Asn Ala Gly Glu Tyr Ala Thr Ala Ala Lys Asn Asn Leu Asn

50 55 60

Ala Phe Ala Glu Ser Leu Lys Thr Glu Ala Ala Val Phe Ser Lys Ser

65 70 75 80

Phe Glu Gly Lys Ala Ser Ala Ser Asp Val Phe Lys Glu Ser Thr Lys

85 90 95

Asn Phe Gln Ala Val Val Asp Thr Tyr Ile Lys Asn Leu Pro Lys Asp

100 105 110

Leu Thr Leu Lys Asp Phe Thr Glu Lys Ser Glu Gln Ala Leu Lys Tyr

115 120 125

Met Val Glu His Gly Thr Glu Ile Thr Lys Lys Ala Gln Gly Asn Thr

130 135 140

Glu Thr Glu Lys Glu Ile Lys Glu Phe Phe Lys Lys Gln Ile Glu Asn

145 150 155 160

Leu Ile Gly Gln Gly Lys Ala Leu Gln Ala Lys Ile Ala Glu Ala Lys

165 170 175

Lys Ala

<210> 173

<211> 311

<212> PROTEIN

<213> Buchnera aphidicola

<400> 173

Met Lys Thr Ser Ser Ser Lys Val Phe Ala Ser Cys Val Ala Ile Val

1 5 10 15

Cys Leu Ala Ser Val Ala Asn Ala Leu Pro Val Gln Lys Ser Val Ala

20 25 30

Ala Thr Thr Glu Asn Pro Ile Val Glu Lys His Gly Cys Arg Ala His

35 40 45

Lys Asn Leu Val Arg Gln Asn Val Val Asp Leu Lys Thr Tyr Asp Ser

50 55 60

Met Leu Ile Thr Asn Glu Val Val Gln Lys Gln Ser Asn Glu Val Gln

65 70 75 80

Ser Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser Glu Gln Ser Asn Glu

85 90 95

Gly Gln Asn Ser Glu Gln Ser Asn Glu Val Gln Ser Ser Glu His Ser

100 105 110

Asn Glu Gly Gln Asn Ser Lys Gln Ser Asn Glu Gly Gln Asn Ser Glu

115 120 125

Gln Ser Asn Glu Val Gln Ser Ser Glu His Ser Asn Glu Gly Gln Asn

130 135 140

Ser Glu Gln Ser Asn Glu Val Gln Ser Ser Glu His Ser Asn Glu Gly

145 150 155 160

Gln Asn Ser Lys Gln Ser Asn Glu Gly Gln Asn Ser Lys Gln Ser Asn

165 170 175

Glu Val Gln Ser Ser Glu His Trp Asn Glu Gly Gln Asn Ser Lys Gln

180 185 190

Ser Asn Glu Asp Gln Asn Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser

195 200 205

Lys Gln Ser Asn Glu Gly Gln Asn Ser Lys Gln Ser Asn Glu Asp Gln

210 215 220

Asn Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser Lys Gln Ser Asn Glu

225 230 235 240

Val Gln Ser Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser Lys Gln Ser

245 250 255

Asn Glu Gly Gln Ser Ser Glu Gln Ser Asn Glu Gly Gln Asn Ser Lys

260 265 270

Gln Ser Asn Glu Val Gln Ser Pro Glu Glu His Tyr Asp Leu Pro Asp

275 280 285

Pro Glu Ser Ser Tyr Glu Ser Glu Glu Thr Lys Gly Ser His Glu Ser

290 295 300

Gly Asp Asp Ser Glu His Arg

305 310

<210> 174

<211> 431

<212> PROTEIN

<213> Buchnera aphidicola

<400> 174

Met Lys Thr Ile Ile Leu Gly Leu Cys Leu Phe Gly Ala Leu Phe Trp

1 5 10 15

Ser Thr Gln Ser Met Pro Val Gly Glu Val Ala Pro Ala Val Pro Ala

20 25 30

Val Pro Ser Glu Ala Val Pro Gln Lys Gln Val Glu Ala Lys Pro Glu

35 40 45

Thr Asn Ala Ala Ser Pro Val Ser Asp Ala Lys Pro Glu Ser Asp Ser

50 55 60

Lys Pro Val Asp Ala Glu Val Lys Pro Thr Val Ser Glu Val Lys Ala

65 70 75 80

Glu Ser Glu Gln Lys Pro Ser Gly Glu Pro Lys Pro Glu Ser Asp Ala

85 90 95

Lys Pro Val Val Ala Ser Glu Ser Lys Pro Glu Ser Asp Pro Lys Pro

100 105 110

Ala Ala Val Val Glu Ser Lys Pro Glu Asn Asp Ala Val Ala Pro Glu

115 120 125

Thr Asn Asn Asp Ala Lys Pro Glu Asn Ala Ala Ala Pro Val Ser Glu

130 135 140

Asn Lys Pro Ala Thr Asp Ala Lys Ala Glu Thr Glu Leu Ile Ala Gln

145 150 155 160

Ala Lys Pro Glu Ser Lys Pro Ala Ser Asp Leu Lys Ala Glu Pro Glu

165 170 175

Ala Ala Lys Pro Asn Ser Glu Val Pro Val Ala Leu Pro Leu Asn Pro

180 185 190

Thr Glu Thr Lys Ala Thr Gln Gln Ser Val Glu Thr Asn Gln Val Glu

195 200 205

Gln Ala Ala Pro Ala Ala Ala Gln Ala Asp Pro Ala Ala Ala Pro Ala

210 215 220

Ala Asp Pro Ala Pro Ala Pro Ala Ala Ala Pro Val Ala Ala Glu Glu

225 230 235 240

Ala Lys Leu Ser Glu Ser Ala Pro Ser Thr Glu Asn Lys Ala Ala Glu

245 250 255

Glu Pro Ser Lys Pro Ala Glu Gln Gln Ser Ala Lys Pro Val Glu Asp

260 265 270

Ala Val Pro Ala Ala Ser Glu Ile Ser Glu Thr Lys Val Ser Pro Ala

275 280 285

Val Pro Ala Val Pro Glu Val Pro Ala Ser Pro Ser Ala Pro Ala Val

290 295 300

Ala Asp Pro Val Ser Ala Pro Glu Ala Glu Lys Asn Ala Glu Pro Ala

305 310 315 320

Lys Ala Ala Asn Ser Ala Glu Pro Ala Val Gln Ser Glu Ala Lys Pro

325 330 335

Ala Glu Asp Ile Gln Lys Ser Gly Ala Val Val Ser Ala Glu Asn Pro

340 345 350

Lys Pro Val Glu Glu Gln Lys Pro Ala Glu Val Ala Lys Pro Ala Glu

355 360 365

Gln Ser Lys Ser Glu Ala Pro Ala Glu Ala Pro Lys Pro Thr Glu Gln

370 375 380

Ser Ala Ala Glu Glu Pro Lys Lys Pro Glu Ser Ala Asn Asp Glu Lys

385 390 395 400

Lys Glu Gln His Ser Val Asn Lys Arg Asp Ala Thr Lys Glu Lys Lys

405 410 415

Pro Thr Asp Ser Ile Met Lys Lys Gln Lys Gln Lys Lys Ala Asn

420 425 430

<210> 175

<211> 160

<212> PROTEIN

<213> Buchnera aphidicola

<400> 175

Met Asn Gly Lys Ile Val Leu Cys Phe Ala Val Val Phe Ile Gly Gln

1 5 10 15

Ala Met Ser Ala Ala Thr Gly Thr Thr Pro Glu Val Glu Asp Ile Lys

20 25 30

Lys Val Ala Glu Gln Met Ser Gln Thr Phe Met Ser Val Ala Asn His

35 40 45

Leu Val Gly Ile Thr Pro Asn Ser Ala Asp Ala Gln Lys Ser Ile Glu

50 55 60

Lys Ile Arg Thr Ile Met Asn Lys Gly Phe Thr Asp Met Glu Thr Glu

65 70 75 80

Ala Asn Lys Met Lys Asp Ile Val Arg Lys Asn Ala Asp Pro Lys Leu

85 90 95

Val Glu Lys Tyr Asp Glu Leu Glu Lys Glu Leu Lys Lys His Leu Ser

100 105 110

Thr Ala Lys Asp Met Phe Glu Asp Lys Val Val Lys Pro Ile Gly Glu

115 120 125

Lys Val Glu Leu Lys Lys Ile Thr Glu Asn Val Ile Lys Thr Thr Lys

130 135 140

Asp Met Glu Ala Thr Met Asn Lys Ala Ile Asp Gly Phe Lys Lys Gln

145 150 155 160

<210> 176

<211> 415

<212> PROTEIN

<213> Buchnera aphidicola

<400> 176

Met His Leu Phe Leu Ala Leu Gly Leu Phe Ile Val Cys Gly Met Val

1 5 10 15

Asp Ala Thr Phe Tyr Asn Pro Arg Ser Gln Thr Phe Asn Gln Leu Met

20 25 30

Glu Arg Arg Gln Arg Ser Ile Pro Ile Pro Tyr Ser Tyr Gly Tyr His

35 40 45

Tyr Asn Pro Ile Glu Pro Ser Ile Asn Val Leu Asp Ser Leu Ser Glu

50 55 60

Gly Leu Asp Ser Arg Ile Asn Thr Phe Lys Pro Ile Tyr Gln Asn Val

65 70 75 80

Lys Met Ser Thr Gln Asp Val Asn Ser Val Pro Arg Thr Gln Tyr Gln

85 90 95

Pro Lys Asn Ser Leu Tyr Asp Ser Glu Tyr Ile Ser Ala Lys Asp Ile

100 105 110

Pro Ser Leu Phe Pro Glu Glu Asp Ser Tyr Asp Tyr Lys Tyr Leu Gly

115 120 125

Ser Pro Leu Asn Lys Tyr Leu Thr Arg Pro Ser Thr Gln Glu Ser Gly

130 135 140

Ile Ala Ile Asn Leu Val Ala Ile Lys Glu Thr Ser Val Phe Asp Tyr

145 150 155 160

Gly Phe Pro Thr Tyr Lys Ser Pro Tyr Ser Ser Asp Ser Val Trp Asn

165 170 175

Phe Gly Ser Lys Ile Pro Asn Thr Val Phe Glu Asp Pro Gln Ser Val

180 185 190

Glu Ser Asp Pro Asn Thr Phe Lys Val Ser Ser Pro Thr Ile Lys Ile

195 200 205

Val Lys Leu Leu Pro Glu Thr Pro Glu Gln Glu Ser Ile Ile Thr Thr

210 215 220

Thr Lys Asn Tyr Glu Leu Asn Tyr Lys Thr Thr Gln Glu Thr Pro Thr

225 230 235 240

Glu Ala Glu Leu Tyr Pro Ile Thr Ser Glu Glu Phe Gln Thr Glu Asp

245 250 255

Glu Trp His Pro Met Val Pro Lys Glu Asn Thr Thr Lys Asp Glu Ser

260 265 270

Ser Phe Ile Thr Thr Glu Glu Pro Leu Thr Glu Asp Lys Ser Asn Ser

275 280 285

Ile Thr Ile Glu Lys Thr Gln Thr Glu Asp Glu Ser Asn Ser Ile Glu

290 295 300

Phe Asn Ser Ile Arg Thr Glu Glu Lys Ser Asn Ser Ile Thr Thr Glu

305 310 315 320

Glu Asn Gln Lys Glu Asp Asp Glu Ser Met Ser Thr Thr Ser Gln Glu

325 330 335

Thr Thr Thr Ala Phe Asn Leu Asn Asp Thr Phe Asp Thr Asn Arg Tyr

340 345 350

Ser Ser Ser His Glu Ser Leu Met Leu Arg Ile Arg Glu Leu Met Lys

355 360 365

Asn Ile Ala Asp Gln Gln Asn Lys Ser Gln Phe Arg Thr Val Asp Asn

370 375 380

Ile Pro Ala Lys Ser Gln Ser Asn Leu Ser Ser Asp Glu Ser Thr Asn

385 390 395 400

Gln Gln Phe Glu Pro Gln Leu Val Asn Gly Ala Asp Thr Tyr Lys

405 410 415

<210> 177

<211> 126

<212> PROTEIN

<213> Sitophilus zeamais

<400> 177

Met Thr Arg Thr Met Leu Phe Leu Ala Cys Val Ala Ala Leu Tyr Val

1 5 10 15

Cys Ile Ser Ala Thr Ala Gly Lys Pro Glu Glu Phe Ala Lys Leu Ser

20 25 30

Asp Glu Ala Pro Ser Asn Asp Gln Ala Met Tyr Glu Ser Ile Gln Arg

35 40 45

Tyr Arg Arg Phe Val Asp Gly Asn Arg Tyr Asn Gly Gly Gln Gln Gln

50 55 60

Gln Gln Gln Pro Lys Gln Trp Glu Val Arg Pro Asp Leu Ser Arg Asp

65 70 75 80

Gln Arg Gly Asn Thr Lys Ala Gln Val Glu Ile Asn Lys Lys Gly Asp

85 90 95

Asn His Asp Ile Asn Ala Gly Trp Gly Lys Asn Ile Asn Gly Pro Asp

100 105 110

Ser His Lys Asp Thr Trp His Val Gly Gly Ser Val Arg Trp

115 120 125

<210> 178

<211> 16

<212> PROTEIN

<213> Acyrthosiphon pisum

<400> 178

Arg Gln Ile Lys Ile Trp Phe Gln Asn Arg Arg Met Lys Trp Lys Lys

1 5 10 15

<210> 179

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Tat Peptide

<400> 179

Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Pro Pro Gln

1 5 10

<210> 180

<211> 18

<212> PROTEIN

<213> Human immunodeficiency virus 1

<400> 180

Leu Leu Ile Ile Leu Arg Arg Arg Ile Arg Lys Gln Ala His Ala His

1 5 10 15

Ser Lys

<210> 181

<211> 27

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Transportan

<400> 181

Gly Trp Thr Leu Asn Ser Ala Gly Tyr Leu Leu Gly Lys Ile Asn Leu

1 5 10 15

Lys Ala Leu Ala Ala Leu Ala Lys Lys Ile Leu

20 25

<210> 182

<211> 27

<212> PROTEIN

<213> Artificial sequence

<220>

<223> MPG

<400> 182

Gly Ala Leu Phe Leu Gly Phe Leu Gly Ala Ala Gly Ser Thr Met Gly

1 5 10 15

Ala Trp Ser Gln Pro Lys Lys Lys Arg Lys Val

20 25

<210> 183

<211> 21

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Pep-1

<400> 183

Lys Glu Thr Trp Trp Glu Thr Trp Trp Thr Glu Trp Ser Gln Pro Lys

1 5 10 15

Lys Lys Arg Lys Val

20

<210> 184

<211> 18

<212> PROTEIN

<213> Artificial sequence

<220>

<223> MAP

<400> 184

Lys Leu Ala Leu Lys Leu Ala Leu Lys Ala Leu Lys Ala Ala Leu Lys

1 5 10 15

Leu Ala

<210> 185

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> R6W3

<400> 185

Arg Arg Trp Trp Arg Arg Trp Arg Arg

15

<210> 186

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Direct primer for Bacillus

<400> 186

gaggtagacg aagcgacctg

20

<210> 187

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Reverse primer for Bacillus

<400> 187

ttccctcacg gtactggttc

20

<210> 188

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Universal primer for bacteria 27F

<400> 188

agagtttgat cmtggctcag

20

<210> 189

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<223> Universal primer for bacteria 1492R

<400> 189

taccttgtta cgactt

16

<210> 190

<211> 1583

<212> DNA

<213> Lactobacillus kunkeei

<400> 190

ttccttagaa aggaggtgat ccagccgcag gttctcctac ggctaccttg ttacgacttc

60

accctaatca tctgtcccac cttagacgac tagctcctaa aaggttaccc catcgtcttt

120

gggtgttaca aactctcatg gtgtgacggg cggtgtgtac aaggcccggg aacgtattca

180

ccgtggcatg ctgatccacg attactagtg attccaactt catgcaggcg agttgcagcc

240

tgcaatccga actgagaatg gctttaagag attagcttga cctcgcggtt tcgcgactcg

300

ttgtaccatc cattgtagca cgtgtgtagc ccagctcata aggggcatga tgatttgacg

360

tcgtccccac cttcctccgg tttatcaccg gcagtctcac tagagtgccc aactaaatgc

420

tggcaactaa taataagggt tgcgctcgtt gcgggactta acccaacatc tcacgacacg

480

agctgacgac aaccatgcac cacctgtcat tctgtccccg aagggaacgc ccaatctctt

540

gggttggcag aagatgtcaa gagctggtaa ggttcttcgc gtagcatcga attaaaccac

600

atgctccacc acttgtgcgg gcccccgtca attcctttga gtttcaacct tgcggtcgta

660

ctccccaggc ggaatactta atgcgttagc tgcggcactg aagggcggaa accctccaac

720

acctagtatt catcgtttac ggcatggact accagggtat ctaatcctgt tcgctaccca

780

tgctttcgag cctcagcgtc agtaacagac cagaaagccg ccttcgccac tggtgttctt

840

ccatatatct acgcatttca ccgctacaca tggagttcca ctttcctctt ctgtactcaa

900

gttttgtagt ttccactgca cttcctcagt tgagctgagg gctttcacag cagacttaca

960

aaaccgcctg cgctcgcttt acgcccaata aatccggaca acgcttgcca cctacgtatt

1020

accgcggctg ctggcacgta gttagccgtg gctttctggt taaataccgt caaagtgtta

1080

acagttactc taacacttgt tcttctttaa caacagagtt ttacgatccg aaaaccttca

1140

tcactcacgc ggcgttgctc catcagactt tcgtccatg tggaagattc cctactgctg

1200

cctcccgtag gagtctgggc cgtgtctcag tcccaatgtg gccgattacc ctctcaggtc

1260

ggctacgtat catcgtcttg gtgggctttt atctcaccaa ctaactaata cggcgcgggt

1320

ccatcccaaa gtgatagcaa agccatcttt caagttggaa ccatgcggtt ccaactaatt

1380

atgcggtatt agcacttgtt tccaaatgtt atcccccgct tcggggcagg ttaccacgt

1440

gttactcacc agttcgccac tcgctccgaa tccaaaaatc atttatgcaa gcataaaatc

1500

aatttgggag aactcgttcg acttgcatgt attaggcacg ccgccagcgt tcgtcctgag

1560

ccaggatcaa actctcatct taa

1583

<210> 191

<211> 1395

<212> DNA

<213> Lactobacillus Firm-4

<400> 191

acgaacgctg gcggcgtgcc taatacatgc aagtcgagcg cgggaagtca gggaagcctt

60

cgggtggaac tggtggaacg agcggcggat gggtgagtaa cacgtaggta acctgcccta

120

aagcggggga taccatctgg aaacaggtgc taataccgca taaacccagc agtcacatga

180

gtgctggttg aaagacggct tcggctgtca ctttaggatg gacctgcggc gtattagcta

240

gttggtggag taacggttca ccaaggcaat gatacgtagc cgacctgaga gggtaatcgg

300

ccacattggg actgagacac ggcccaaact cctacgggag gcagcagtag ggaatcttcc

360

acaatggacg caagtctgat ggagcaacgc cgcgtggatg aagaaggtct tcggatcgta

420

aaatcctgtt gttgaagaag aacggttgtg agagtaactg ctcataacgt gacggtaatc

480

aaccagaaag tcacggctaa ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg

540

ttgtccggat ttattgggcg taaagggagc gcaggcggtc ttttaagtct gaatgtgaaa

600

gccctcagct taactgagga agagcatcgg aaactgagag acttgagtgc agaagaggag

660

agtggaactc catgtgtagc ggtgaaatgc gtagatatat ggaagaacac cagtggcgaa

720

ggcggctctc tggtctgtta ctgacgctga ggctcgaaag catgggtagc gaacaggatt

780

agataccctg gtagtccatg ccgtaaacga tgagtgctaa gtgttgggag gtttccgcct

840

ctcagtgctg cagctaacgc attaagcact ccgcctgggg agtacgaccg caaggttgaa

900

actcaaagga attgacgggg gcccgcacaa gcggtggagc atgtggttta attcgaagca

960

acgcgaagaa ccttaccagg tcttgacatc tcctgcaagc ctaagagatt aggggttccc

1020

ttcggggaca ggaagacagg tggtgcatgg ttgtcgtcag ctcgtgtcgt gagatgttgg

1080

gttaagtccc gcaacgagcg caacccttgt tactagttgc cagcattaag ttgggcactc

1140

tagtgagact gccggtgaca aaccggagga aggtggggac gacgtcaaat catcatgccc

1200

cttatgacct gggctacaca cgtgctacaa tggatggtac aatgagaagc gaactcgcga

1260

ggggaagctg atctctgaaa accattctca gttcggattg caggctgcaa ctcgcctgca

1320

tgaagctgga atcgctagta atcgcggatc agcatgccgc ggtgaatacg ttcccgggcc

1380

ttgtacacac cgccc

1395

<210> 192

<211> 220

<212> PROTEIN

<213> Sitophilus zeamais

<400> 192

Met Lys Glu Thr Thr Val Val Trp Ala Lys Leu Phe Leu Ile Leu Ile

1 5 10 15

Ile Leu Ala Lys Pro Leu Gly Leu Lys Ala Val Asn Glu Cys Lys Arg

20 25 30

Leu Gly Asn Asn Ser Cys Arg Ser His Gly Glu Cys Cys Ser Gly Phe

35 40 45

Cys Phe Ile Glu Pro Gly Trp Ala Leu Gly Val Cys Lys Arg Leu Gly

50 55 60

Thr Pro Lys Lys Ser Asp Asp Ser Asn Asn Gly Lys Asn Ile Glu Lys

65 70 75 80

Asn Asn Gly Val His Glu Arg Ile Asp Asp Val Phe Glu Arg Gly Val

85 90 95

Cys Ser Tyr Tyr Lys Gly Pro Ser Ile Thr Ala Asn Gly Asp Val Phe

100 105 110

Asp Glu Asn Glu Met Thr Ala Ala His Arg Thr Leu Pro Phe Asn Thr

115 120 125

Met Val Lys Val Glu Gly Met Gly Thr Ser Val Val Val Lys Ile Asn

130 135 140

Asp Arg Lys Thr Ala Ala Asp Gly Lys Val Met Leu Leu Ser Arg Ala

145 150 155 160

Ala Ala Glu Ser Leu Asn Ile Asp Glu Asn Thr Gly Pro Val Gln Cys

165 170 175

Gln Leu Lys Phe Val Leu Asp Gly Ser Gly Cys Thr Pro Asp Tyr Gly

180 185 190

Asp Thr Cys Val Leu His His Glu Cys Cys Ser Gln Asn Cys Phe Arg

195 200 205

Glu Met Phe Ser Asp Lys Gly Phe Cys Leu Pro Lys

210 215 220

<210> 193

<211> 13

<212> PROTEIN

<213> Pandinus imperator

<400> 193

Phe Leu Ser Thr Ile Trp Asn Gly Ile Lys Gly Leu Leu

1 5 10

<210> 194

<211> 13

<212> PROTEIN

<213> Urodacus yaschenkoi

<400> 194

Ile Leu Ser Ala Ile Trp Ser Gly Ile Lys Ser Leu Phe

1 5 10

<210> 195

<211> 13

<212> PROTEIN

<213> Scorpiops tibetanus

<400> 195

Leu Trp Gly Lys Leu Trp Glu Gly Val Lys Ser Leu Ile

1 5 10

<210> 196

<211> 22

<212> PROTEIN

<213> Apostichopus japonicus

<400> 196

Phe Pro Phe Leu Lys Leu Ser Leu Lys Ile Pro Lys Ser Ala Ile Lys

1 5 10 15

Ser Ala Ile Lys Arg Leu

20

<210> 197

<211> 13

<212> PROTEIN

<213> Urodacus yaschenkoi

<400> 197

Ile Leu Ser Ala Ile Trp Ser Gly Ile Lys Gly Leu Leu

1 5 10

<210> 198

<211> 27

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Uy192 + cell penetrating peptide

<400> 198

Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Phe Leu Ser Thr Ile

1 5 10 15

Trp Asn Gly Ile Lys Gly Leu Leu Phe Ala Met

20 25

<---

Claims (10)

1. A method of increasing the adaptability of a honey bee to organophosphate insecticides, the method comprising: administering to the honey bee a composition containing an effective amount of bacteria that metabolize organophosphate insecticides, formulated with a carrier edible for insects, wherein the bacteria that metabolize organophosphate insecticides are selected from the group consisting from Snodgrassella spp., Bartonella spp., Parasaccharibacter spp. and Lactobacillus spp., and are able to colonize the bee’s intestines, thereby increasing its fitness. 2. The method of claim 1, wherein administering the composition comprises delivering the composition to a honey bee hive or at least one habitat where a honey bee grows, lives, reproduces or feeds. 3. Method according to any one of paragraphs. 1, 2, where the composition is a liquid, solid, aerosol, paste, gel or gas. 4. Method according to any one of paragraphs. 1-3, wherein the organophosphate insecticide is fenitrothion. 5. Method according to any one of paragraphs. 1-4, where the carrier is a means for coating the seeds. 6. Method according to any one of paragraphs. 1-5, where a honey bee is found in a honey bee colony. 7. A composition for increasing the adaptability of a honey bee to organophosphorus insecticides, containing an effective amount of bacteria that metabolize organophosphorus insecticides, formulated with an insect-edible carrier in the form of a liquid, solid, aerosol, paste, gel or gas, where the carrier is a seed coating and wherein the organophosphate insecticide metabolizing bacteria are selected from the group consisting of Snodgrassella spp., Bartonella spp., Parasaccharibacter spp. and Lactobacillus spp., and are able to colonize the bee intestines. 8. The composition according to claim 7, wherein bacteria that metabolize organophosphorus insecticides metabolize fenitrothion. 9. The composition according to claim 7 or 8, wherein the bacteria that metabolize organophosphorus insecticides are present in a concentration of at least 100,000 cells/ml. 10. Method according to any one of paragraphs. 1-6, in which bacteria that metabolize organophosphorus insecticides are present in a concentration of at least 100,000 cells/ml.

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