DK2021362T3 - Fluorescent proteins and genes encoding them - Google Patents

Description

Description [0001] The present invention relates to a class of new fluorescent proteins and genes encoding them.

Background [0002] Fluorescent proteins (FPs), in particular green fluorescent proteins, are commonly used fluorescent makers in molecular biology to monitor gene expression and protein localization in living organisms and in medical diagnostic applications.

[0003] Fluorescent proteins are found in a variety of marine organisms ranging from the jellyfish Aequorea victoria, to the Indo-Pacific coral Discosoma. WO 2004/058973 discloses fluorescent proteins isolated from Copepoda species, and genes encoding said proteins. The proteins are used for e.g. labelling of biomolecules. Due to their genetically encoded fluorescence, fluorescent proteins have become most important marker molecules and tools in cell biology. Becoming spontaneously fluorescent without any requirements for cofactors, substrates or other gene products, FPs have revolutionized research in many areas of biology.

[0004] During recent years FPs have also gathered strong appreciation as powerful tools for the drug discovery process. As fluorescent probes, FPs enable both real-time and non-invasive reporting in living cells. This ability provides a basis for cell-based monitoring of FP-linked targets upon administration of external drugs. The impact of FPs has been revolutionary; FPs have not only facilitated visualization of intricate cellular architecture but they have also acted as markers of protein dynamics and behavior in cell biology. These applications have been translated to drug discovery where fluorescence proteins have been utilized in fluorescence and confocal imaging, HTS/HCS screening assays and for in vivo diagnostics. FPs cannot only be used in early stage target characterization but also in retrieving non-invasive 'whole organism' data and in evaluating lead compound toxicology.

[0005] Limitations of most fluorescent proteins are generally associated with molecular brightness and/or stability. Moreover, many FPs have additional complications involving protein folding, chromophore maturation and self-association. Although FPs have vastly improved over the years, mainly by introducing mutations, they still exhibit major limitations.

[0006] There is an interest in obtaining new fluorescent proteins with different properties compared to known fluorescent proteins. For instance, there are no fluorescent proteins on the market that can be used in paraffin sections at room temperature for immunohistochemical purposes since they lose their fluorescence intensity under such conditions.

Description of the invention [0007] The present invention provides a class of new fluorescent proteins with different properties compared to known proteins, e.g. they can be used in paraffin sections at room temperature for immunohistochemical purposes since they retain their fluorescence intensity under such conditions.

[0008] One aspect of the invention is directed to an isolated and purified structural gene encoding a fluorescent protein encoding a mutant or recombinant protein that has at least 80% amino acid sequence identity with the fluorescent protein, and comprising the internal amino acid sequence

Gly Tyr Xaa Xaa Xaa Gin Tyr Leu Pro Xaa Pro (SEQ ID NO: 47) 1 5 10 wherein

Xaa in position 3 is Ala or Gly,

Xaa in position 4 is Phe, His or Tyr,

Xaa in position 5 is His, Tyr or Asn, or Xaa in position 10 is Phe or Tyr.

[0009] The term "structural gene" means the protein coding nucleotide sequence of a gene or polynucleotide.

[0010] The internal sequence SEQ ID NO:47 is found in all hitherto analyzed proteins of the new class of fluorescent proteins, but some amino acid substitution, extension and/or deletion in this sequence may be possible, especially in the positions where there are variations in the amino acids, i.e. where the amino acid is Xaa.

[0011] A mutant or recombinant protein that has at least 80% amino acid sequence identity with a fluorescent protein defined in this invention may be truncated and/or have amino acid substitutions, insertions and/or deletions and have any percentage of amino acid identity with regard to the fluorescent protein defined in this invention between 80% and 99.9 %, such as at least 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9 % identity.

[0012] In an embodiment of the invention the structural gene is selected from the group consisting of SEQ ID NOs: 1-23.

[0013] In another embodiment of the invention the structural gene is selected from the group consisting of SEQ ID NOs: 48 - 67.

[0014] In still another embodiment of the invention the structural gene is selected from the group consisting of SEQ ID NOs: 88 - 90.

[0015] The nucleotide sequences SEQ ID NOs: 88 -90 are examples of isolated and purified structural genes encoding mutant proteins of the invention. The exemplified mutant sequences have the sequence of the wild-type Branchiostoma lanceolatum nucleotide sequence SEQ ID NO: 59 from position 7 including position 666, and some point mutations. The nucleotide A in positions 356 and 357 of the mutant sequences SEQ ID NO: 88-90, respectively, has been inserted instead of the nucleotide C in the wild-type sequence, and further the nucleotide C in position 524 of the mutant sequences SEQ ID NOs: 89 and 90, respectively, has been inserted instead of the nucleotide A in the wild-type sequence. In addition, the mutant sequence SEQ ID NO: 90 has in position 469 the nucleotide A instead of G in the wild-type sequence and in position 471 the nucleotide G instead of C in the wild-type sequence.

[0016] Another aspect of the invention is directed to a vector comprising a structural gene according to the invention, such as a structural gene selected from the group consisting of SEQ ID NOs: 1-23, SEQ ID NOs: 48 - 67 and SEQ ID NOs: 88 -90. The vector may be any vector which can comprise a structural gene of the invention and necessary flanking regions with regulatory elements necessary for expression of the desired fluorescent protein of the invention or a fusion protein comprising such a protein according to the invention. The regulatory elements necessary for expression are e.g. a suitable operon or promoter that is natural or foreign to the host selected for expression of the protein. Suitable vectors useful in the present invention are e.g. plasmids, cosmids and virus expression vectors.

[0017] Yet another aspect of the invention is directed to a host cell comprising a vector according to the invention or comprising a transgene including a structural gene according to the invention. The transgene should be operably inserted into the genome of the host to express the desired fluorescent protein of the invention or a fusion protein comprising such a protein according to the invention. Suitable host cells are both prokaryotic cells such as Escherichia coli cells, and eukaryotic cells such as mammalian, insect, yeast, and plant cells.

[0018] The internal sequence SEQ ID NO:47 may possibly have some amino acid substitution, extension and/or deletion in this sequence, especially in the positions where there are variations in the amino acids, i.e. where the amino acid is Xaa.

[0019] In an embodiment of the invention the protein of the invention is an isolated or recombinant fluorescent protein or an isolated mutant or recombinant protein that has at least 80% amino acid sequence identity with the fluorescent protein, and has the Internal amino acid sequence

Gly Tyr Xaa Xaa Xaa Gin Tyr Leu Pro Xaa Pro (SEQ ID NO: 47) 1 5 10 wherein

Xaa in position 3 is Ala or Gly,

Xaa in position 4 is Phe, His or Tyr,

Xaa in position 5 is His, Tyr or Asn, or Xaa in position 10 is Phe or Tyr.

[0020] In analyzed wild-type proteins according to the Invention, the C-terminal Pro in SEQ ID NO: 47 is followed by Asp Gly, Ala Gly, Asp Asp or Gly Gly.

[0021] A mutant or recombinant protein that has at least 80% amino acid sequence identity with a fluorescent protein defined in this invention may be truncated and/or have amino acid substitutions, insertions and/or deletions and have any percentage of amino acid identity with regard to the fluorescent protein defined in this invention between 80% and 99.9 %, such as at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9 % identity.

[0022] In the invention the fluorescent protein is a protein that has an amino acid sequence selected from the group consisting of SEQ ID NOs: 24 -46, SEQ ID NOs: 68 -87, and SEQ ID NOs: 91-93.

[0023] The amino acid sequences SEQ ID NOs: 91 -93 are examples of mutant proteins of the invention that have the amino acid sequence SEQ ID NO: 79 of the wild-type Branchiostoma lanceolatum and some point mutations. In the amino acid sequences SEQ ID NOs: 91 -93, the Thr of the wild-type protein in position 119 has been replaced by Lys, and further in the sequences SEQ ID NOs: 92 and 93, the amino acid Asn of the wild-type protein in position 175 has been replaced by Thr. Additionally, the amino acid Asp of the wild-type protein in position 157 has been replaced by Lys in the sequence SEQ ID NO: 93.

[0024] The recombinant fluorescent proteins comprised by the present invention may be in monomeric, dimeric or multimeric, such as tetrameric, form.

Use of fluorescent proteins according to the invention [0025] Due to the inherent property of FPs to spontaneously become fluorescent, and in all organisms and in all types of cells, FPs have become invaluable tools in many biological and medical disciplines. A wide range of applications of the FPs have been developed that can be classified into four areas; visualizing / monitoring of organisms, cells, proteins and molecular events. The fluorescent proteins and structural genes encoding the them according to the invention may be used in all hitherto known applications of fluorescent proteins, such as those described below.

[0026] To visualize an organism, a structural gene encoding a FP can be introduced and together with appropriate regulatory sequences become expressed as an inheritable fluorescent marker in a variety of organisms ranging from virus, bacteria and yeast to plants, fish and mice. For example, infections of viruses and bacteria can be monitored, as well as the survival and spread of genetically modified organisms, GMOs. Since the FPs according to the invention can be functionally expressed in both prokaryotic and eukaryotic cells, and they have excellent stability, brightness and photoresistance, they are expected to be excellent for such applications.

[0027] To visualize cells or organelles, a structural gene encoding a FP may be introduced as a transgene in e.g. germ line cells and in vitro cultured cells. For example, monitoring of cell fate/lineages in transgenic animals, of cancer cells in vivo, of wound healing and of neurite outgrowth can be accomplished. Additional examples are marking of organelles (mitochondria, nuclei; etc) and GFP imaging: methodology and application to investigate cellular compartmentation in plants. (See e.g. for a review J Exp Bot. 2001 Apr;52(356):529-39.) The properties of the FPs according to the invention enables them to be used for paraffin-imbedded and section tissues.

[0028] To visualize proteins, a structural gene encoding a FP can be fused to a gene of interest producing a fusion protein that is tagged by the FP chromophore. The fusion protein can then be monitored in e.g. living cells in real time, thus enabling analyzes of cellular localization of individual proteins (numerous examples in the prior art).

[0029] Protein-protein interactions can be followed by labeling two different proteins with two different chromophores, and their interaction can be monitored by FRET (Fluorescence resonance energy transfer) or BRET (Bioluminescence Resonance Energy Transfer) in case of a bioluminescent donor to a fluorescent acceptor protein.

[0030] In drug screening protein-drug interactions are studied. Co-localization of fluorescent fusion proteins with intracellular localization markers are used as indicators of movements of intracellular fluorescent fusion proteins/peptides. The aggregation or internalization of fluorescent-tagged plasma membrane proteins (e.g. G-protein coupled receptors) can be used as drug screening assays.

[0031] In the literature there are numerous references to the use of FPs and genes encoding them as sensors for different purposes, such as sensors for protease activity: Detection of MMP activity in living cells by a genetically encoded surface-displayed FRET sensor. Biochim Biophys Acta 2007 Man;1773(3):400-7, Epub 2006 Nov 11, and Development and application of a GFP-FRET intracellular caspase assay for drug screening. J Biomol Screen. 2000 Oct,5(5):307-18;as sensors for atoms or ions: Genetic oxygen sensor: GFP as an indicator of intracellular oxygenation Adv Exp Med Biol. 2005;566:39-44, Elimination of environmental sensitivity in a cameleon FRET-based calcium sensor via replacement of the acceptor with Venus. Cell Calcium. 2005

Apr,37(4) :341 -8, Construction of a whole-cell gene reporter for the fluorescent bioassay of nitrate. Anal Biochem. 2004 May 1 ;328(1 ):60-6, and Transgenic mice expressing a pH and Cl- sensing yellow-fluorescent protein under the control of a potassium channel promoter. Eur J Neurosci. 2002 Jan;15(1 ):40-50/as sensors for organic molecules: A new green fluorescent protein-based bacterial biosensor for analyzing phenanthrene fluxes. Environ Microbiol. 2006 Apr;8(4):697-708, and Live imaging of glucose homeostasis in nuclei of COS-7 cells. J Fluoresc. 2004 Sep;14(5):603-9/as sensors for electrical activity or neural cell activation: A hybrid approach to measuring electrical activity in genetically specified neurons". Nat Neurosci. 2005 Nov;B(11):1619-26. Epub 2005 Oct, and A genetically encoded optical probe of membrane voltage. Neuron. 1997 Oct;19(4):735-41 :as sensors for cell cycle: Characterization and gene expression profiling of a stable cell line expressing a cell cycle GFP sensor. Cell Cycle. 2005 Jan;4(1 ):191 -5. Epub 2005 Jan 29; as sensors for promoters or gene activation; A high-throughput approach to promoter study using green fluorescent protein. Biotechnol Prog. 2004 Nov-Dec;20(6):1634-40; and as sensors for apoptosis: Degradation of GFP-labelled POM121, a non-invasive sensor of nuclear apoptosis, precedes clustering of nuclear pores and externalization of phosphatidylserine. Apoptosis. 2004 May;9(3):363-8.

[0032] The invention will now be illustrated by description of drawings and of embodiments and experiments of the invention, but it should be understood that the scope of for the invention is not limited to any described details.

Short description of the drawings [0033]

Fig-1 is a diagram that shows emission spectra after excitation at 500-520 nm of four FPs of the invention, namely Green Y-1 = SEQ ID NO: 68 ; Yellow 0-1 = SEQ ID NO: 74; Orange 0-2 = SEQ ID NO: 75 and Red R-5 = SEQ ID NO: 79.

Fig. 2 is a diagram that shows the fluorescence plotted against time for the reference EGFP and two FPs of the invention, Green FP Y-1 = SEQ ID NO: 68;

Red FP R-4 = SEQ ID NO: 78, indicating that the FPs are resistant to photobleaching.

Fig. 3 is a diagram that shows the fluorescence plotted against pH for the reference EGFP and a red FP, R-4 = SEQ ID NO: 78, of the invention, indicating that R-4 is insensitive to pH.

Fig. 4 is a diagram that shows the fluorescence plotted against temperature for the reference EGFP (squares) and Red FP (triangles), R-1= SEQ ID NO: 76, and Green FP (circles), Y-3 = SEQ ID NO: 70, of the invention. The fluorescence was recorded after 1 h (and 1 min for EGFP) after incubation at the indicated temperature.

Fig. 5 is a diagram that shows the fluorescence plotted against concentration of ethanol for the reference EGFP (squares) and Red FP (triangles), R-1= SEQ ID NO: 76, and Green FP (circles), Y-3 = SEQ ID NO: 70, of the invention. The fluorescence was recorded after 1 h after incubation at the indicated concentration of ethanol.

Fig. 6 is a diagram that shows absorbance plotted against elution volume in size exclusion chromatography of the wild-type FP wt R-5 (SEQ ID NO: 79), the double mutant Mut B (SEQ ID NO: 92) and the triple mutant Mut C (SEQ ID NO: 93) as well as EGFP.

Fig. 7 is a diagram that shows normalized fluorescence plotted against wavelength in excitation and emission spectra of mutant R-5C, SEQ ID NO: 93. The emission spectra were recorded after excitation at 590 nm.

Description of embodiments and experiments of the invention

Natural occurrence of fluorescent proteins of the invention [0034] Proteins comprised by the definition of fluorescent proteins according to the invention occur naturally in the Cephalochordata (amphioxus) e.g. Branchiostoma floridae or 8ranchiostoma Janceolatum. They are expressed in supportive cells in the anteriormost of the body, e.g., cells in the coelom walls, the subepidermal canals, the oral cirri skeleton, and the oral cirri tufts. However, the number of different positive cell types varies between different individuals.

[0035] The proteins are found as either only a single fluorescent protein, or as a mixture of different fluorescent proteins.

[0036] All properties have been examined at room temperature unless otherwise stated. Characteristics of the fluorescent proteins of the invention [0037] The fluorescence characteristics of selected FPs have been determined by confocal laser scanning microscopy on bacterial colonies with samples in 96-well plates and on proteins purified after expression in E. coli. Brightness was calculated as the product of quantum yield and molar extinction coefficient-determined by comparing the Coomassie Brilliant Blue staining intensity, after SDS-PAGE, of known amounts of EGFP (Enhanced Green Fluorescence Protein) with the new FPs. Purified EGFP has been employed as reference fluorescent protein.

Optical properties of fluorescent proteins of the invention [0038] The purified proteins are yellow to orange in solution, and yellow to red in solid state.

[0039] The absorption maxima for the purified native proteins are at 210 nm (peptide bonds), 260 nm (aromatic amino acid residues), and 484-490 nm (fluorophore). Upon denaturation, the peak at 260 nm is shifted to 280 nm, and the fluorophore peak is almost completely lost.

[0040] The emission spectrum of fluorescent cells in situ and of purified proteins consists of peaks at 485, 500, 515, 530, 545, 560, 575, 590, 610 and 635 nm. The number of peaks varies between individuals and between different cell types in the same individual. The absorption maxima for each individual protein might be slightly shifted from these values due to overlaying of neighbouring peaks.

[0041] Excitation at 458 nm results in peaks at 485 and 500 nm (primary excitation), and 545-635 nm (presumably fluorescence resonance energy transfer, FRET) Excitation at 476 or 486 nm results in peaks at 515 and 530 nm (primary excitation), and 545-635 nm (presumably FRET). Excitation at 514 nm results in peaks at 545-635 nm (primary excitation).

[0042] The excitation spectra of all analyzed new FPs are very similar with maxima around 510-520 nm, in accordance with the identity of the chromophore forming residues, but in spite of this similarity the emission spectra differ considerable as shown in Figure 1. This property thus suggests a new mechanism for generation of colour diversity as compared to other known fluorescent proteins.

Resistance to bleaching [0043] The fluorescence has similar bleaching resistance in strong light as the most resistant fluorescent protein (EGFP (Enhanced Green Fluorescence Protein) and Emerald). The photostability, i.e. resistance to bleaching, was determined on proteins in solutions (Tris-HCI, pH 7.5) that were placed between a cover slip and a microscope slide and exposed to the highest possible light intensity in a fluorescent microscope (mercury lamp, 100X oil immersion). As shown in Figure 2, the new FPs are very resistant to bleaching and display an even greater photostability than EGFP.

Maturation and Stoke's shift [0044] As other red fluorescent proteins, the new red variants go through a maturation stage before reaching the mature form that emits red light. In contrast to many other wild-type red FPs the maturation step is quite fast as red fluorescence can be observed in growing E. coli colonies.

[0045] Another interesting feature of the red variants is the very large difference between absorption and emission max, the Stoke's shift is more than 100 nm.

Insensitivity to environmental effects [0046] The fluorescence is pH dependent. Fluorescence occurs in weakly acidic, neutral and basic solutions but not in acidic and strongly basic solutions. The green fluorescence has a maximum at pH 10, at least 50% fluorescence between pKa1 and pKa2 (limits for >50% of maximal fluorescence; pKa1 = 7.7, and pKa2 = 11.6), and has almost a linear dependence of pH between pH 6-9.

[0047] The new FPs are extremely stable and insensitive to environmental effects. The pH sensitivity has been analyzed with a red variant (R-4 = SEQ ID NO: 78) and as shown in Figure 3, the fluorescence of R-4 is maintained over a wider pH range than the fluorescence of EGFP.

Physical properties of fluorescent proteins of the invention [0048] The apparent molecular weight of a native, proteinase K-treated protein is about 25 kDa (gel filtration) and of denaturated protein is about 30-35 kDa (SDS-PAGE). The green fluorescent proteins are slightly larger than the red ones. The protein oligomerizes and predominantly forms tetramers. However, trimers and pentamers, hexamers, dimers and octamers, and polymers are also present (in decreasing order of frequency).

Chemical properties of fluorescent proteins of the invention [0049] The proteins are soluble in water, phosphate buffer and Tris buffer but not in acetone, ethanol, glycerol or xylene. The proteins can be precipitated with acetone or ethanol but not with ammonium sulfate (80% of saturated solution).

[0050] The fluorescence is lost upon denaturation.

[0051] The fluorescence resists proteinase K (0.1-1 mg/ml at 45°C overnight), detergents (10% sodium dodecyl sulfate, and 0.1% triton X-100), aldehyde fixation (formaldehyde, and glutaraldehyde), chelates (1 M EDTA), many organic solvents (acetone, ethanol, glycerol, melted paraffin, and xylene), high salinity (80% saturated ammonium sulfate, 4M sodium chloride, and saturated disodium hydrogen phosphate), low salinity (distilled water), heavy metal ions (cupper chloride, lead nitrate, and silver nitrate), weak oxidizing agents (hydrogen peroxide, oxygen in air, potassium chromate in neutral solution, potassium dichromate in neutral solution, potassium ferricyanide, silver nitrate, and sodium chlorate in neutral solution), weak reducing agents (10 mM dithiothreitol, and pyrogallol in neutral solution), and moderately high temperatures (45°C for 12 h, 65°C for 1 h).

[0052] The fluorescence is destroyed by some organic solvents e.g. (benzyl alcohol -benzyl benzoate mixture, strong oxidizing agents (iodine, periodic acid potassium chromate in acid solution, potassium dichromate in acid solution, potassium permanganate, sodium chlorate in alkaline solution, and sodium hypochlorite in alkaline solution), strong reducing agents (pyrogallol in alkaline solution), and very high temperature (98°C).

Stability of the fluorescent proteins [0053] The stability of the FPs has been analyzed both in situ and using purified proteins after expression in E. coli. As shown in Figures 4 and 5, two of the recombinantly expressed proteins of the invention, Red FP (R-1= SEQ ID NO: 76) and Green FP (Y-3 = SEQ ID NO: 70), are very thermostable and withstand high concentrations of ethanol. The new FPs are also stable in e.g. 6 M guanidine hydrochloride, and withstand many organic solvents making them useful in histochemical applications including those using paraffin imbedded and sectioned tissues.

Bright red FPs [0054] The brightness (product of quantum yield and molar extinction coefficient) has been calculated for two of the new red variants, and they are clearly among the brightest red wild-type proteins ever found. The Table below summarizes the fluorescence characteristics of selected members of the FPs of the invention.

Table . Fluorescence characteristics of selected FPs or the invention and a selection of reference proteins (from Ref. 1).

Isolating fluorescent proteins from Cephalochordata [0055] Several hundred specimens of lancelet, i.e. Cephalochordata, also named amphioxus, are collected. In this example Branchiostoma lanceolatum were collected. Their heads were cut off and mixed with an equal volume of a neutral buffer solution (e.g., 10 mM Tris, pH 7.5,10 mM NaCI). The mixture was digested with proteinase K (final concentration of 0.1 mg/ml) at 40°C over-night followed by centrifugation for 10 min at 16 000 rpm to remove any remaining debris. The supernatant was loaded on a Sephadex G-200 column (30-100 cm; equilibrated with the same buffer as used during the digestion) and gel filtrated. Fluorescent fractions were collected, pooled, and precipitated by addition of 1.8 volumes of acetone and centrifugation for 1 min at 16000 rpm. The supernatant was discharged. The pellet was washed with 65% acetone, briefly dried (allowing remaining acetone to evaporate), dissolved in water, denaturated with sodium dodecyl sulfate (SDS; final concentration of 1 %) and dithiothreitol (DTT; final concentration 100 mM) at 95°C for 3 min, and loaded on a 2.5% SDS-PAGE gel. After the completed gel electrophoresis and Coomassie staining, the two bands at around 30 kDalton were cut out, eluted, and digested into fragments with trypsin. The fragments are analyzed with tandem mass spectrometry using MALDI-TOF (matrix assisted laser desorption-time of flight) to obtain their amino acid sequence. Degenerated oligonucleotide primers are designed from these amino acid sequences. These primers are used for 5'-RACE PCR (rapid amplification of cDNA ends polymerase chain reaction) on cDNA that is prepared from purified mRNA from lancelet heads. The PCR products are size separated by agarose electrophoresis. The different bands are cut out, purified, cloned, and sequenced. Oligonucleotide primers are designed from the obtained sequences and used for 3'RACE PCR on the same cDNA to obtain the complete coding region. The PCR products are cloned into an expression vector, and fluorescent transformants are selected and sequenced. All steps are performed at roomtemperature unless otherwise stated.

Isolating fluorescent proteins from genome project [0056] The known amino acid sequence of green fluorescent proteins (GFP) from copepods (obtained from GenBank) are used for searches through the unassembled trace files obtained from genomic sequencing of Florida lancelet (Branchiostoma floridae) (available at http://www.ensembl.org/). Nucleotide sequences containing putative GFP-like exons were selected. These sequences were elongated by successive searches with the sequences for new matching sequences and alignments of these new sequences to these already found. This is repeated for many cycles until either complete genes were obtained, or no more new sequences are found. The assembled genes are analyzed by a splicing prediction software [NetGene2 software (http://www.cbs.dtu.dk/services/NetGene2/)] and putative exons are converted into amino acid sequences. The obtained nucleotide sequences are analyzed for conserved regions. Degenerated oligonucleotide primers are designed for two different conserved regions (located in exon 3 and exon 6). These primers are used for nested 5'-RACE PCR on cDNA that is prepared from purified mRNA from lancelet heads. The PCR products are size separated by agarose electrophoresis. The different bands are cut out, purified, cloned, and sequenced. Oligonucleotide primers are designed from the obtained sequences and used for 3'RACE PCR on the same cDNA to obtain the complete coding region. The PCR products are cloned into an expression vector, and fluorescent transformants are selected and sequenced.

Cloning and sequencing [0057] RNA was prepared and through RT-PCR, using degenerate PCR -primers, and 5'-and 3'-RACE, full-length cDNA clones could subsequently be obtained by fluorescence screening of E. coli colonies. The obtained full-length clones represent yellow/green, orange and red FPs. No blue or pure green FPs were among the full-length clones, but a number of incomplete clones were sequenced that may represent these colours.

[0058] In total some 40 unique sequences encoding novel fluorescent proteins have been obtained so far. The individual proteins are structurally closely related and although distinct from other known fluorescent proteins they clearly belong to the same superfamily.

Mutant proteins a. Aggregation state [0059] The wild-type fluorescent proteins have a tendency to self-aggregate and form tetramers. Among the limited number of new FPs that we so far have analyzed, no monomeric variant has been observed, but the tendency to self-aggregate appears to vary among individual variants. In order to generate a pure monomeric variant, which would be advantageous for certain applications, the protein R-5 (SEQ ID NO: 79) was subjected to limited mutagenesis after extensive 3-D modelling. Three amino acid residues were chosen as suitable candidates for mutagenesis, and three mutants with single, double and triple mutations were constructed and custom made by an external laboratory. After expression in E. coli and purification their aggregation state were analyzed by gel-chromatography on a HiPrep Sephacryl S-200 column. The conditions used (i.e. high protein concentrations) were chosen to promote aggregation, and EGFP, which is a weak forming dimer, did elute as a peak with an apparent molecular weight of -60 kDa, i.e. as a dimer. As shown in Figure 6, the double mutant of R-5 (Mut B), SEQ ID NO: 92, also eluted as a dimer whereas the triple mutant (Mut C), SEQ ID NO: 93, eluted with an apparent molecular weight of -30 kDa, i.e. as a monomer. b. Characteristics of mutant fluorescent proteins [0060] The dimeric mutant (Mut B), SEQ ID NO: 92, has a diminished tendency to selfaggregate, but folds less efficiently than the wild-type protein. The optical properties appear to be the same as the wild-type R-5 (SEQ ID NO: 79) protein. Neither excitation-emission spectra, nor brightness or stability appear different from the wild-type protein. However, the monomeric mutant (Mut C), SEQ ID NO: 93, has lost much of the brightness of the wild-type protein, and - probably due to a maturation defect - has its major emission peak shifted to the orange part of the spectra. Interestingly, the mutants, including the monomeric variant, have also a new emission peak with a maximum at -660 nm with excitation at 590 nm. The emission peak is quite wide and although the overall intensity is low, there is a clear emission up to 750 nm, thus extending into the infrared part of the spectra (Figure 7), which is a clear benefit for in vivo applications.

SEQUENCE LISTING

[0061]

<110> Innoventus Project AB <120> Fluorescent proteins and genes encoding them <130> P40600048PCT00 <150> US 60/811,769 <151 >2006-06-08 <150> SE 0601261-1 <151 >2006-06-08 <160> 93 <170> Patentln version 3.4 <210> 1 <211 >480

<212> DNA <213> Branchiostoma floridae <400> 1 atgcctcttc caactaccca cgagttgcac atctttggtt ccttcaatgg tgtggagttt 60 gacttggtgg gccgtggcga aggtaatcca aaggatgggt ctcagaacct acacctgaag 120 tccaccaagg gtgccctcca gttctcccca tggatgctgg tccctcacat cgggtacggc 180 ttctaccagt acctgcctta cccggacggc gaaatgtcgc cttaccaggc cgccatgtat 240 ggtggctcag ggtacctaat gcatcgcaca atgcaatatg aagacggtgc caagattagt 300 ggccactaca aatacaccta cgagggaagc cacgtgaaag gagaatttca gctcattggg 360 accggattcc ctactgacgg tcctgtgatg accaaccagc tcaccgctgc ggactggtgc 420 gtggacaagt tgctgtaccc caacgacaag accattatca gcaagttcga ctggagctac 480

<210> 2 <211 >678 <212> DNA <213> Branchiostoma floridae <400>2 atggcttcca tattggtgcc tcttccaaag acccacgagt tgcacatctt tggttccttc 60 aatggtgtta agtttgacat ggttggtgaa ggcaccggta accctaatga ggggtctgag 120 gagctaaaac tgaagtccac caatggtccc ctgaagttct ccccctacat cctggtccct 180 cacctcgggt acgctttcaa ccagtacctg cccttccctg acgggatgtc gcctttccag 240 gccgccatgc aggacgaatc ggggtatcaa gtgcatcgca cgttgcagta tgaagacggt 300 gccttcgtga ctgctaactt acgctacacc tacgagggaa gtcacatcaa aggggagttc 360 caggtgatcg ggaccggttt tcctcctgat ggtcctgtga tgaccaacaa gctcaccgct 420 ttggactgga gcgtggtcaa gtttgtgtac cccaacgaca agaccatcct cagcactttc 480 gacaaaacct acaccaccac cgatggcaaa cgctaccagt gcacatttcg tgaaaacaac 540 accttcgcca agccgatggc ggccgacatc ctgcagaagc agccgatgtt cattttccat 600 aagacggagc tccagcactc taacaacgcc gagctcacct tcaaggagaa gcagacagcc 660 ttctccgata tgaagtga 678

<210> 3 <211 >479 <212> DNA <213> Branchiostoma floridae <400>3 atgtctctgc ctacggccca tgactgccac atgttcggct ccatcaacgg ccacgagttt 60 gacctggtgg gcggtggaaa cgggaacccg aatgacggga cactggagac caaggtgcgc 120 tccaccaagg gtgccctgcc cttctccccc gtgatcctgg cccctaacct ggggtacggg 180 taccaccagt acctgccctt cccggccggg acctcaccgt accagcaggc catcaccaac 240 ggagtgtacc aaaagcaccg caccttcaag ttcgaggacg gcggcgtcat gaccatcaac 300 ttccgctaca cctactcagg gaacaagatc aagggagagt tccacgtact ggttggatrcc 360 gggttccctg atgacggccc tgtgaagacc cactcactcc agcagcatga tcataacgtg 420 gagaggctga tggtgctggg agacaagacc atcggcagcg acaacatgtg gactttcac 479

<210> 4 <211 >473 <212> DNA <213> Branchiostoma floridae <400>4 tctctgccta cgacccatga ctgccacatg ttcggctcca tcaacggcca cgagtttgac 60 ctggtgggcg gtggaaacgg gaacccgaat gacgggacgc tggagaccaa ggtgcgctcc 120 accaagggag ccctgccctt ctccccagtg atcctggccc ctaacctggg gtacgggtac 180 caccagtacc tgcccttccc ggccgggacc tcaccctacc agcaggccat caccaacgga 240 gggtaccaaa agcaccgcac gttcaagttc gaggacggcg gcgtcatgac catcaacttc 300 cgctacacct actcagggaa caagatcaag ggagagttcc acgtggttgg atccgggttc 360 ccagatgatg gccctgtgat gaccaactct ctccagcagc aegatcataa cgtggagagg 420 ctgatggtgc tgggagacaa gaccatcggc agcgacaaca tgtggacttt cac 473

<210> 5 <211 >672 <212> DNA <213> Branchiostoma floridae <400>5 atgcctcttc caaagaccca cgagttgcac atctttggtt ccttcaatgg tgttgagttt 60 gacatggttg ctcgcggcat cggcaaccct aatgaggggt ctgaggaact gaacgcgaag 120 ttcaccaagg gtcccctgaa gttctccccc tacatcctgg tccctcacct cgggtacgct 180 tactaccagt acctgccctt ccctgacggg atgtcgcctt tccaggccgc catgcacgac 240 ggctcgggct atcaagtgca tcgcacgatt cagtatgaag acggtgcctc cgtgactgcc 300 cactaccgct acacctacga gggaagccac atcaaagggg agtttcaggt gatcgggacc 360 ggatttcctc ctgacggtcc tgtgatgacc aacaagctca ccgctatgga ctggagcgtg 420 accaagatgc tgtacccgaa cgacaagacc atcctcagca ctgccgactg tagctacacc 480 accaccgagg gcaaacgcta ccagagcaaa atgcgtgaaa acaacacctt cgccaagccg 540 atggcggccg acatcctgca gaagcagccg atgttcgtgt tccgtaagac ggagctccag 600 cactccaaga ccgagctcac cttcaaggag tggcagaaag ccttcaccga tgtgataact 660 ggacatatct aa 672

<210> 6 <211 >696 <212> DNA <213> Branchiostoma floridae <400>6 atgtctgtcc cgacaaacct cgacttgcac atctacggct ccatcaacgg tatggagttc 60 gacatggtgg gcggtgggag tggcaaccca aaggatggat cgctggccgt aaacgtgaag 120 tccaccaaag gagctctgtg cgtttccccc ctgctggtgg gcccgcatct ggggtacggc 180 cactaccagt acctaccctt ccctgacggc gcgtcgcctt tccaggcagc cgtgaacaac 240 ggcgggtatc aaatgcatcg cagtttcaac ttcgaggacg gggccgtgct gactgccacc 300 tacaactact cctacagcgg cggcaagatc cagggagagt ttcatctggt gggcagcggt 360 ttccccgacg atagtccggt gatgaccaac gcgctgaccg gtctggacag gagcgtgtcc 420 aagctgatgt gcacgtccga tgacaagctc gtcgagtccg tgcactggag ctaccgcacc 480 agcagcggcg ggcgctaccg tgccacggtg cagaccaact tcaccttcgc aaagcccatc 540 gaagctggcc tgaagaacaa catgccgatg ttcgtgttcc gtcagctgga agtcaccggc 600 tccaaaaccg agatcggcct tcaggagcag caaaaggcgt tctccaccgt tctgatgcgt 660 ttatggttga aatgcaaaag agttgaaata ctttga 696

<210> 7 <211 >693 <212> DNA <213> Branchiostoma floridae <400>7 tctgtcccga caaacctcga cttgcacatc tacggctcca tcaacggtat ggagttcgac 60 atggtgggcg gtggcagtgg caacccaaag gatggatcgc tggccgtaaa cgtgaagtcc 120 accaaaggag cactgcgcgt ttcccccctg ctggtgggcc cgcatctggg gtacggccac 180 taccagtacc tacccttccc cgacggcgcg tcgcctttcc aggcagccgt gaacaacggc 240 gggtatcaaa tgcatcgcag tttcaacttc gaggacgggg ccgtgctgac tgccacctac 300 aactactcct acagcggcgg caagatccag ggagagtttc atctggtggg cagcggtttc 360 cccgacgata gtccggtgat gaccaacgcg ctgaccggtc tggacaggag cgtgtccaag 420 ctgatgtgca cgtccgatga caagctcgtc gagtccgtgc actggagcta ccgcaccagc 480 agcggcgggc gctaccgtgc cacggtgcag accaacttca ccttcgcaaa gcccatcgca 540 gctggcctga agaacaacat gccgatgttc gtgttccgtc agctggaagt caccggctcc 600 aaaaccgaga tcggccttca ggagcagcaa aaggcgttct ccaccgttct gatgcgttta 660 tggttgaaat gcaaaagagt tgaaatactt tga 693

<210> 8 <211 >699 <212> DNA <213> Branchiostoma floridae <400>8 atgtctgtcc cgacaaacct cgacttgcac atctacggct ccatcaacgg tatggagttt 60 gacatggtgg gcggtgggag tggcaaccca aatgacggat cgctgagcgt aaacgtgaag 120 tctaccaaag gtgccctgcg cgtttctcct ctgctggtgg gcccgcatct ggggtacggc 180 cactaccagt acctaccctt ccctgacggt ccgtcgcctt tccaggcagc cgtgaacaac 240 ggcggatatc aaatgcatcg ctctttcaac ttcgaggacg gtgccgtgct gactgccacc 300 tacaactact cctacagcgg cggcaagatc cagggagagt ttcatgtact ggtgggcagc 360 tgtttccccg acgatagtcc ggtgatgacc aacgcgctga ccggtttgga caggagcgtg 420 gccaagctga tgtgcgtgtc cgatgacaag cttgccgagt tcgtggactg gacctaccgc 480 accagcagcg gcgggcgcta ccgtgccacg gtgcagacca acttcacctt cgcaaagccc 540 atcgcagctg gcctgaagaa caacatgccg atgttcgtgt tccgtcagct ggaagtcacc 600 ggctccaaaa ccgagatcag ccttcaggag cagcaaaagg cgttctccac cgttctggtg 660 cgtttatggt tgaaatgtaa aagagctgaa atactttga 699

<210> 9 <211 >660 <212> DNA <213> Branchiostoma floridae <400>9 atgcctcttc caaagaccca cgagttgcac atctttggtt ccttcaatgg tgttgagttt 60 gacatggttg gtcgcggcat cggcaaccct aatgaggggt ctgaggaact gaacgcgaag 120 ttcaccaagg gtcccctgaa gttctccccc tacatcctgg tcccccacct cgggtacgct 180 tactaccagt acctgccctt ccctgacggg atgtcgcctt tccaggccgc catgcacgac 240 ggctcgggct atcaagtgca tcgcacgatt cagtatgaag acggtgcctc cgtgactgcc 300 cactaccgct acacctacga gggaagccac atcaaagggg agtttcaggt gatcgggacc 360 ggatttcctc ctgacggtcc tgtgatgacc aacaagctca ccgctatgga ctggagcgtg 420 accaagatgc tgtacccgaa cgacaagacc atcctcagca ctgtcgactg tagctacacc 480 accaccgagg gcaaacgcta ccagagcaaa atgcgtgaaa acaacacctt cgccaagccg 540 atggcggccg acatcctgca gaagcagccg atgttcgtgt tccgtaagac ggagctccag 600 cactccaaga ccgagctcac cttcaaggag tggcagaaag ccttcaccga tgtgatgtga 660

<210> 10 <211> 663 <212> DNA <213> Branchiostoma floridae <400> 10 atgcctcttc caaagaccca cgagttgcac atctttggtt ccttcaatgg tgttaagttt 60 gacatggttg gtgaaggcac cggtaaccct aatgaggggt ctgaggagct aaaactgaag 120 tccaccaatg gtcccctgaa gttctccccc tacatcctgg tccctcacct cgggtacgct 180 ttcaaccagt acctgccctt ccctgacggg atgtcgcctt tccaggccgc catgcaggac 240 gaatcggggt accaagtgca tcgcacgttg cagtatgaag acggtgcctt cgtgactgct 300 aacttacgct acacctacga gggaagtcac atcaaagggg agttccaggt gatcgggacc 360 ggttttcctc ctgatggtcc tgtgatgacc aacaagctca ccgctttgga ctggagcgtg 420 gtcaagtttg tgtaccccaa cgacaagacc atcctcagca ctttcgacaa aacctacacc 480 accaccgagg gcaaacgcta ccagtgcaca tttcgtgaaa acagcacctt cgccaagccg 540 atggcggccg acatcctgca gaagcagccg atgttcatct tccataagac ggagctccag 600 cactccaaca atgccgagct caccttcaag gagaagcaga cagctttctc cgatatgaag 660 tga 663

<210> 11 <211 >669 <212> DNA <213> Branchiostoma floridae <400> 11 cctcttccaa cgactcacga ggtgcacgtc tatggctcta tcaacggtgt tgagtttgac 60 ttggtgggta gcggcaaagg caacccgaag gatggttctg aggagatcca agtgaagtcc 120 actaagggtc ccctcgggtt ctccccgccc gtcgtggtcc ccaacatcgg gtacggcttc 180 caccagtact tgcccttccc cgacgggatg tcgcctttcc aggccgccgc ggacgatggc 240, tcggggtacg tagtccatcg taatattcag tttgaagacg gcgcctcgct gactggcatc 300 taccgatatt cctacgatgc aggtcacatc aaaggagagt ttcgtgtggt tggcagtggt 360 ttccctgctg acggtcctgt gatgaccaaa tcgctcacgg ctgtggactg gagcgtggct 420 accatgctgt tcccgaacga caccaccgtt gtctccacca ttgactggac ttgccccact 480 accagcggca aacgctacca cgccacggtg aggaccaact acaccttcgc caagccgata 540 gcgggcagca ttctccagaa gcagccaatg ttcgtgtttc gtaagacgga agtcaaggcc 600 tctgactccg agatcaacct caaggagtcg cagaaggcct ttcatgacct cgttggtata 660 tgtatttga 669

<210> 12 <211 >654 <212> DNA <213> Branchiostoma floridae <400> 12 atgcctctgc ccgcaaccca tgaaatccac cttcacggct ctgtcaacgg ccacgagttc 60 gacttggtgg gcagtggaaa aggtgacccg aaagccggct cgctggtgac cgaagtgaaa 120 tccaccatgg gtcgcctgaa gttctctcct cacttgatga tcccccacct cgggtacggg 180 tactaccagt acctccccta cccggacgga ccctcgcctt tccagaccgc catgctcgat 240 ggatcggggt ataaagtcca ccgtgtgttc aactttgagg acggtggcgt gttgtccatc 300 gactacaatt atgcctacga ggggactcac atcaagtccg actttaagct gatgggaagc 360 ggtttccctg acgacggccc agtcatgacc agccaaattg tcgaccagga cggctgcgtg 420 tccaagaaga cgtatcttaa cgacaacacc atcgtggaca gcttcgactg gtcttacaat 480 ctgcagaatg ggaagcgcta cagggctcga gtgacgagca actacatctt cgggaagccc 540 ctcgcggccg atgtaatgaa gaagcagccg gtcttcgtgt accgcaagtg ctacgtgaag 600 tctacccaga ccgagatcac cctggacgag agggagaagg cgttctacga agtg 654

<210> 13 <211 >696 <212> DNA <213> Branchiostoma floridae <400> 13 atgtctgttc cgacaaacct cgacctgcac atctatggct ccatcaacgg tatggagttc 60 gacatggtgg gcggtgggag tggcaaccca aaggacggat cgctgagcgt aaatgtgaag 120 tcaaccaaag gtgccctgcg cgtttctcct ctgctggtgg gcccgcatct ggggtacggc 180 cactaccagt acctaccctt ccctgacgat ccgtcgcctt tccaggcagc cgtgaacaac 240 ggcgggaatc aaatgcatcg ctctttcaac ttcgaggacg gtgccgtgct gactgccacc 300 tacaactact cctacagcgg cggcaagatc cagggagagt ttcatctggt gggcagctgt 360 ttccccaacg atagtccggt gatgaccaac gcgctgaccg gtctggacag gagcgtggcc 420 aagctgatgt gcgtgtccga tgacaagctt gccgagttcg tggactggac ctaccgcacc 480 agcagcggcg ggcgctaccg tgccacggtg cagaccaact tcaccttcgc aaagcccatc 540 gcagctggcc tgaagaacaa catgccgatg ttcgtgttcc gtcagctgga agtcaccggc 600 tccaaaaccg agatcggcct tcaggagcag caaaaggcgt tctccaccgt tctggtgcgt 660 tcatggttga aatataaaag agctgaaata ctttga 696

<210> 14 <211 >657 <212> DNA <213> Branchiostoma floridae <400> 14 atgtctctcc ctacgactca cgaccttcac atcttcggct ecgtcaatgg cgcggagttc 60 gacctggtgg gaggcggaaa gggcaacccg aacgatggaa cgctggagac cagtgtgaaa 120 tccacccggg gcgccctgcc ctgctccccg ctgctgatcg gacccaacct ggggtacggc 180 ttctaccagt acctgccctt ccctggcggc gcctcaccct tccaaaccgc catcacggac 240 ggagggtacc aggttcaccg tgtgttcaag tttgaagacg gcggagtgct gagttgcaac 300 ttccgctaca cctacgaggg cggcaagatc aaaggggagt tccagctgat cgggtcaggt 360 ttccctgccg gcgggcctgt gatgtccggc ggactgacca ccctggacag gagcgtggcc 420 aaactgcagt gctcggacga ccgcaccatc accggcacta acaactggag cttctgcacc 480 accgatggga agcgccacca ggcggatgtg cagacgaact acaccttcgc caagccgctc 540 ccggccggtc tgaaggagaa gatgccgatc ttcctggggc accagatcga ggtcaaggcg 600 tccaagaccg agatcaccct gagcgagaaa gtgaaggcct tcatcgacac tgtgtaa 657

<210> 15 <211 >762 <212> DNA <213> Branchiostoma floridae <400> 15 atgcacgcta caaaacacac tgtaggttgt tatccactgt atctagggca ccagtctctc 60 cctaccactc acgaccttca catcttcggc tccgtcaatg gcgcggagtt cgacctggtg 120 ggaggcggaa agggcaaccc gaacgatgga acgctggaga ccagtgtgaa atccacccgg 180 ggcgccctgc cctgctcccc gctgctgatc ggacccaacc tggggtacgg cttctaccag 240 tacctgccct tccctggcgg cgcctcaccc ttccaaaccg ccatcacgga cggagggtac 300 caggttcacc gtgtgttcaa gtttgaagac ggcggagtgc tgaattgcaa cttccgcaac 360 ttccgctaca cctacgaggg cggcaagatc aaaggggagt tccagctgat cgggtcaggt 420 ttccctgccg gcgggcctgt gatgcccggc ggactgacca ccctggacag gagcgtggcc 480 aaactgcagt gctcggacga ccgcaccatc accggcacta acaactggag cttctgcacc 540 accgatggga agcgccacca ggcggatgtg cagacgaact acacttcgcc aagccgctcc 600 ggccggtctc aaggagaaga tgccggtctt cctggggcac cagatcgagg tcaaagcgtc 660 caagaccgag atcaccctga gcgagaaagt gaaggccttc atcgacactg tgtgaagttc 720 aagttcgccg actgtgttaa gcccagaatt cagtcctgtt aa 762

<210> 16 <211 >668 <212> DNA <213> Branchiostoma floridae <400> 16 tctctcccaa cggctcacga ccttcacatc ttcggctccg tcaatggcgc ggagttcgac 60 ctggtgggag gcggaaaggg caacccgaac gatggaacgc tggagaccag tgtgaaatcc 120 acccggggcg ccctgccctg ctccccgctg ctgatcggac ccaacctggg gtacggctrc 180 taccagtacc tgcccttccc tggcggcgcc tcacccttcc aaaccgccat cacggacgga 240 ggttaccagg trcaccgtgt gttcaagttt gaagacggcg gagtgctgaa ttgcaacttc. 300 cgctacacct acgagggcgg caagatcaaa ggggagttcc agctgatcgg gtcaggtttc 360 cctgccggcg gtcctgtgat gtccggcgga ctgaccaccc tggacaggag cgtggccaaa 420 ctgcagtgct cggacgactg caccatcacc ggcactaaca actggagctt ctgcaccacc 480 gatgggaagc gccatcaggc ggatgtgcag acgaactaca tcttcgccaa gccgctcccg 540 gccggtctga aggagaagat gccgatcttc ctggggcacc agatcgaggt caaggcgtcc 600 aagaccgaga tcaccctgtg cgagaaagtg aaggccttca tcgacactgt gatctgcttg 660 agatttag 668

<210> 17 <211> 669 <212> DNA <213> Branchiostoma floridae <400> 17 atgagtccaa tttcacctct tccaacgact cacgaggtgc acgtctatgg ctctatcaac 60 ggtgttgagt ttgacttggt gggtagcggc aaaggcaacc cgaaggatgg ttctgaggag 120 atccaagtga agtccactaa gggtcccctc ggattctccc cgtacatcgt ggtgcccaac 180 atcgggtacg gcttccacca gtacctgccc ttcccagacg ggatgtcgcc tttccaggcc 240 gctgcggacg atggctcggg ctacgtagtc catcgtacga ttcagtttga agacggtgcc 300 tcgctgactg gcaactaccg atattcctac gatggaggcc acatcaaagg agagtttcat 360 gtggttggca gcggtttcct tggtgacggc cctgtgatga ccaaatcgct cacggctgtg 420 gactggagtg tggcgaccat gctcttccca aacgacacca ccgttgtctc caccattgac 480 tggacttgcc ccactaccag cggcaaacgc taccatgcca cggtgaggac caactacacc 540 ttcgccaagc cgatagcggc cagcattctc cagaagcagc cgatgttcgt gttccgtaag 600 acggaagtta aggcctctga cgcggagatc aaccttaaga gtggcagaag gctttccatg 660 acctgtgag 669

<210> 18 <211 >666 <212> DNA <213> Branchiostoma floridae <400> 18 atgcctctcc caacgaccca cgaattgcac atctttggtt ccttcaatgg tgtggagttt 60 gatatggttg gtcgcggcat cggcaaccct aatgacgggt atgaggagct aaacctgaag 120 tccaccaagg gtgccctcaa gttctccccc tggatcctgg tgcctcagat cgggtacggc 180 ttccaccagt acctgcctta cccggacgga atgtcgcctt tccaggccgc catgcaggac 240 ggctcagggt accaagtcca tcgcacgatg cagtttgaag acggtgcctc tctgactgcc 300 cacttccgct acacctacga gggaagccac atcaaagggg agtttcaggt gatcgggacc 360 ggattccctg ctgacggtcc tgtgatgacc aacaagctca ccgctgcgga ctggtgcgtg 420 gtcaagatgg tgtaccccaa cgacaagacc atcctcagca ctttcgactg gacctacacc 480 accaccgagg gcaaacgcta ccagagtacg gtgaggacca actacacctt cgccaagccg 540 atggccgcca acatcctgca gaagcagccg atgttcgtgt tccgtaagac ggagctccag 600 • cactccaaga ccgagcttac cttcaaggag tggcagaaag ccttcaccga tgtgatggtg 660 ttttaa 666

<210> 19 <211 >660 <212> DNA <213> Branchiostoma floridae <400> 19 atgcctctgc ccgcaaccca tgagatccac atctatggct ccgtcaacgg ccacgagttc 60 gacctggtgg gcggagggaa aggcgacccg aacgccggct cgctggtgac cgaagtaaaa 120 tccaccatgg gtcccctgaa gttctctccc cacttgatga tcccccacct cgggtacggg 180 tactaccagt acctccccta cccggacgga ccatcgcctt tccagaccgc catgctcgat 240 ggatcggggt attcagtcca tcgcgtgttc gacttcgaag acggaggcaa gctgaccctc 300 gagtttaagt actcctacga gggttcccat atcaaggccg acatgaagtt cacgggaagc 360 ggtttccctg acgacggtcc agtcatgacc agccagattg tcgacgaaga cggctgcgtg 420 tctaagaaca ccatccataa cgacaacacc atcgtggaca acttcgactg gactaatgtc 480 ctacagaatg gaaagcgcta cagggcccac gtgaccagcc actacatctt cgggaagccc 540 ctcgcggccg atgtaatgaa gaagcagccg gtcttcgtgt accgcaagtg ctacgtgaag 600 tctaccaaga ccgagatcac cctggacgag cgagagaagg cgttctacga ggtggtttag 660

<210> 20 <211 >660 <212> DNA <213> Branchiostoma floridae <400> 20 atgcctctgc ccgcaactca tgaaatccac ctccacggct ccatcaacgg ccacgagttc 60 gacctggctg gcggtggaaa aggcgacccg aacgccggct cgctggtgac cacagcgaaa 120 tccaccaagg gtcccctgaa gttctctccc cacttgatga tcccccacct cgggtacggg 180 tactaccagt acctccccta cccggacgga ccctcgcctt tccaggccac catgttggaa 240 ggatcggggt atacagtcca ccgcgtgttt gacttcgaag atggaggcaa gctgtccatc 300 gagtttaagt'actcctacga gggttcccat attaaggccg acatgaagtt cacgggaacc 360 ggtttccctg aggatgggcc ggtcatgacc agccagattg tcgaccagga cggctgcgtg 420 tccaagaaca cctacctcaa cgacaacacc atcgtggaca acttcgactg gacttacaat 480 ctgcagaacg gaaagcgcta cagagcccga gtgacgagcc actacatctt cgacaagccc 540 ttttcagccg atctcatgaa gaagcagccg gtcttcgtgt accgcaagtg ccacgtgaag 600 gcttccaaga ccgagatcaa cctcgacgag agggagaagg cgttctatga gtcggcttga 660

<210> 21 <211> 660 <212> DNA <213> Branchiostoma floridae <400> 21 atgcctctgc ccgcaaccca cgacatccac cttcåcggct ccatcaacgg ccacgagttc 60 gacatggtgg ggggaggaaa aggcgacccg aacgccggct cgctggtgac cacagcgaaa 120 tccaccaagg gtgccctgaa gttctctccc tacttgatga tcccccacct cgggtacggg 180 tactaccagt acctccccta cccggacgga ccctcgcctt tccagacctc catgttggaa 240 ggatcggggt atgcagtcta ccgcgtgttc gactttgaag acggaggcaa gctgactacc , 300 gagtttaagt actcctacga gggttcccat atcaaggccg acatgaagct gatgggaagc 360 ggtttccctg acgacggccc agtcatgacc agccagattg tcgaccagga cggctgcgtg 420 tccaagaaga cgtatcttaa caacaacacc atcgtggaca gcttcgactg gagttacaac 480 ctgcagaatg ggaagcgcta cagggcccga gtgtcgagcc actacatctt cgacaagccc 540 ttttcagccg atctcatgaa gaagcagccg gtcttcgtgt accgcaagtg ccacgtgaag 600 gcttccaaga ccgaagtcac cctggacgag agggagaagg cgttctacga gctggcttag 660

<210> 22 <211> 660 <212> DNA <213> Branchiostoma floridae <400> 22 atgcctctgc ccgcaaccca tgaaatccac cttcåcggct ccgtcaacgg ccacgagttc 60 gacttggtgg gcggtggaaa aggcgacccg aaagccggct cgctggtgac cgaagtgaaa 120 tccaccaagg gtcccctgaa gttttctccc cacttgatga tcccccacct cgggtacggg 180 tactaccagt acctccccta cccggacgga ccctcgcctt tccagaccgc catgctcgat 240 ggatcggggt acaaagtcca ccgtgtgttc aactttgagg acggtggcgt gttgtccatc 300 gagtacaatt attcctacga gggaactcac atcaagtccg actttaagct gatgggaagc 360 ggtttccctg acgacggccc agtcatgacc agccagattg tcgaccagga cggctgcgtg 420 tccaagaaga cgtatcttaa cgacaacacc atcgtggaca gcttcgactg gtcttacaac 480 ctacagaatg ggaagcgcta cagggcccga gtgacgagca actacatctt cgggaagcct 540 ctcgctgccg atgttatgaa gaagcagccg gtcttcgttt accgcaagtg ttacgtgaag 600 tctaccaaga ccgagatcac cctggacgag agggagaagg cgttctatga gctggcttag 660

<210> 23 <211 >657 <212> DNA <213> Branchiostoma floridae <400> 23 cctcttccaa caacccacga gttgcatatc tttggtacct tcaatggtgt cgagtatgac 60 atggtgggcc gtggcaaggg taaccctaat gatggatacg aagagctaaa cctgaagtcc 120 accaagggtc ccctcaagtt ctccccatgg atcctggtcc ctcagattgg gtacggcttc 180 caccagtacc tgccctaccc tgacgggatg tcgcctttcc aggccgccat gcacgatggc 240

tccggctatc aagtgcatcg cacgctggac tttgaagacg gtgccaccct gactgccgac BOO ttccgctaca cctacgaggg gagccacatc aaaggagagt ttaaggtgat cgggaccgga 360 ttccctgctg acggtcctgt gatgaccaac aagctcactg ctgcggactg gtgtgtgaac 420 aagatgctgt acccggacga caagaccatc aacagcacct tcgactggag ctacaccact 480 tccgagggca aacgctacca gagcacagtg agggaaaact acaccttcgc caagccaatg 540 gccgccaaca tcctgcagaa gcagccgatg ttcgtgttcc gtaagacgga gctccagcac 600 tccaagaccg agctcacctt caaggagtgg cagaaagcct tcaccgatgt gatgtaa 657

<210> 24 <211 >220 <212> PRT <213> Branchiostoma floridae <400> 24

Met Pro Leu Pro Thr Thr His Glu Leu His Ile Phe Gly Ser Phe Asn 15 10 15

Gly val Glu Phe Asp Leu Val Gly Arg Gly Glu Gly Asn Pro Lys Asp 20 25 30

Gly ser Gin Asn Leu His Leu Lys Ser Thr Lys Gly Ala Leu Gin Phe 35 40 45 ser Pro Trp Met Leu Val Pro His Ile Gly Tyr Gly Phe Tyr Gin Tyr 50 55 60

Leu pro Tyr pro Asp Gly Glu Met Ser Pro Tyr Gin Ala Ala Met Tyr 65 70 75 80

Gly Gly ser Gly Tyr Leu Met His Arg Thr Met Gin Tyr Glu Asp Gly 85 90 95

Ala Lys ile ser Gly m's Tyr Lys Tyr Thr Tyr Glu Gly ser His val 100 105 U0

Lys Gly Giu Phe Gin Leu Ile Gly Thr Gly phe Pro Thr Asp Gly pro 115 120 125

Val Met Thr Asn Gin Leu Thr Ala Ala Asp Trp cys val Asp Lys Leu 130 135 140

Leu Tyr pro Asn Asp Lys Thr ile ile ser Lys Phe Asp Trp ser Tyr 145 150 155 160

Thr Thr Thr Asp Gly Lys Arg Tyr Gin Ala Lys val Gin Thr Asn Phe 165 170 175

Asp Phe Ala Lys pro Met Ala Ala Asn Tyr Leu Gin Lys Gin Pro Met 180 185 190

Phe val phe Arg Lys Thr Glu Leu Gin His ser Lys Thr Glu Leu Lys 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe His Asp ile Met 210 215 220

<210> 25 <211 >228 <212> PRT <213> Branchiostoma floridae <400> 25

Met Ala ser Ile Leu Val Pro Leu Pro Lys Thr His Glu Leu His ile 15 10 15

Phe Gly Ser Phe Asn Gly val Lys Phe Asp Met val Gly Glu Gly Thr 20 25 30

Gly Asn Pro Asn Glu Gly ser Glu Glu Leu Lys Leu Lys ser Thr Asn 35 40 45

Gly Pro Leu Lys Phe ser Pro Tyr ile Leu val pro His Leu Gly Tyr 50 55 60

Ala Phe Asn Gin Tyr Leu Pro Phe Pro Asp Gly Met Ser Pro Phe Gin 65 70 75 80

Ala Ala Met Gin Asp Glu ser Gly Tyr Glu Asp Gly Ala Phe Val Thr 85 90 95

Ala Asn Leu Arg Tyr Thr Tyr Glu Gly Ser His Ile Lys Gly Glu Phe 100 105 110

Gin His Tyr Asn Val Asp Tyr Asn ile Ile Leu Gin val ile Gly Thr 115 120 125

Gly Phe Pro Pro Asp Gly Pro val Met Thr Asn Lys Leu Thr Ala Leu 130 135 140

Asp Trp ser val val Lys Phe val Tyr Pro Asn Asp Lys Thr Ile Leu 145 150 155 * 160 ser Thr Phe Asp Lys Thr Tyr Thr Thr Thr Asp Gly Lys Arg Tyr Gin 165 170 175

Cys Thr Phe Arg Glu Asn Asn Thr Phe Ala Lys Pro Met Ala Ala asp 180 185 190

Ile Leu Gin Lys Gin Pro Met Phe ile Phe His Lys Thr Glu Leu Gin 195 200 205

His ser Asn Asn Ala Glu Leu Thr Phe Lys Glu Lys Gin Thr Ala Phe 210 215 220

Ser Asp Met Lys 225

<210> 26 <211 > 164 <212> PRT <213> Branchiostoma floridae <400> 26

Met Ser Leu Pro Thr Ala His Asp cys His Met Phe Gly ser ile Asn 15 10 15

Gly His Glu Phe Asp Leu val Gly Gly Gly Asn Gly Asn Pro Asn Asp 20 25 30

Gly Thr Leu Glu Thr Lys Val Arg Ser Thr Lys Gly Ala Leu Pro Phe 35 40 45

Ser Pro val Ile Leu Ala pro Asn Leu Gly Tyr Gly Tyr His Gin Tyr 50 55 60

Leu pro Phe pro Ala Gly Thr ser Pro Tyr Gin Gin Ala ile Thr Asn 65 70 75 80

Gly val Pro ser cys ser Arg Tyr Gin Lys His Arg Thr Phe Lys Phe 85 90 95

Glu Asp Gly Gly Val Met Thr Ile Asn Phe Arg Tyr Thr Tyr ser Gly 100 105 110

Asn Lys ile Lys Gly Glu Phe His val Leu Val Gly ser Gly phe Pro 115. 120 125

Asp Asp Gly Pro val Lys Thr His ser Leu Gin Gin His Asp His Asn 130 135 140 val Glu Arg Leu Met val Leu Gly Asp Lys Thr ile Gly ser Asp Asn 145 150 155 160

Met Trp Thr Phe

<210> 27 <211> 150 <212> PRT <213> Branchiostoma floridae <400> 27

Ser Leu Pro Thr Thr His Asp cys His Met Phe Gly ser Ile Asn Gly 1 5 10 15

His Glu Phe Asp Leu Val Gly Gly Gly Asn Gly Asn Pro Asn Asp Gly 20 .25 30

Thr Leu Glu Thr Lys val Arg ser Thr Lys Gly Ala Leu pro phe ser 35 40 45 pro val Ile Leu Ala pro Asn Leu Gly Tyr Gly Tyr His Gin Tyr Leu 50 55 60 pro Phe Pro Ala Gly Thr ser Pro Tyr Gin Gin Ala ile Thr Asn Gly 65-70 75 80

Gly Phe Glu Asp Gly Gly val Met Thr ile Asn Phe Arg Tyr Thr Tyr 85 90 95 ser Gly Asn Lys Ile Lys Gly Glu Phe His val Val Val Gly ser Gly 100 105 110

Phe Pro Asp Asp Gly Pro Val Met Thr Asn Ser Leu Gin Gin His asd 115 120 125

His Asn val Glu Arg Leu Met val Leu Gly Asp Lys Thr ile Gly ser 130 135 140

Asp Asn Met Trp Thr Phe 145 150

<210> 28 <211 > 216 <212> PRT <213> Branchiostoma floridae <400> 28

Met Pro Leu Pro Lys Thr His Glu Leu His ile Phe Gly ser Phe Asn 15 10 15

Gly val Glu Phe Asp Met val Ala Arg Gly Ile Gly Asn pro Asn Glu 20 25 30

Gly ser Glu Glu Leu Asn Ala Lys Phe Thr Lys Gly Pro Leu Lys Phe 35 40 45 *

Ser Pro Tyr Ile Leu val pro His Leu Gly Tyr Ala Tyr Tyr Gin Tvr 50 55 60 y

Leu Pro Phe Pro Asp Gly Met ser Pro Phe Gin Ala Ala Met His Asd 65 70 75 80

Gly ser Gly Tyr Glu Asp Gly Ala ser val Thr Ala His Tyr Arq Tvr 85 90 95

Thr Tyr Glu Gig ser His ile Lys Gig Glu Phe Gin val ne Gly Thr

Gly Phe Pro Pro Asp Gly Pro val Met Thr Asn Lys Leu Thr Ala Met 115 120 125

Asp Trp ser Val Thr Lys Met Leu Tyr Pro Asn Asp Lys Thr ile Leu 130 135 140 ser Thr Ala Asp Cys ser Tyr Thr Thr Thr Glu Gly Lys Arq Tvr Gin 145 150 155 y ΥΓ 160

Ser Lys Met Arg Glu Asn Asn Thr Phe Ala Lys Pro Met Ala Ala asd 165 170 175

Ile Leu Gin Lys Gin Pro Met Phe val Phe Arg Lys Thr Glu Leu Gin 180 185 190

His ser Lys Thr Glu Leu Thr Phe Lys Glu Trp Gin Lys Ala Phe Thr 195 200 205

Asp val Met ile Thr Gly His ile 210 215

<210> 29 <211> 236 <212> PRT <213> Branchiostoma floridae <400> 29

Met ser val Pro Thr Asn Leu Asp Leu His ile Tyr Gly ser ile Asn 15 10 15

Gly Met Glu Phe Asp Met Val Gly Gly Gly ser Gly Asn Pro Lys Asp 20 25 30

Ser Thr Lys Gly Ala Leu Cys Val Ser Pro Leu Leu Val Gly Pro His 35 40 45

Leu Gly Tyr Gly His Tyr Gin Tyr Leu Pro Phe Pro Asp Gly Ala Ser 50 55 60

Pro Phe Gin Ala Ala val Asn Asn Gly Gly Tyr Gin Met His Arg ser 65 70 75 80 phe Asn Phe Glu Asp Gly Ala val Leu Thr Ala Thr Tyr Asn Tyr ser 85 90 95

Tyr ser Gly Gly Lys ile Gin Gly Glu Phe His Lys Glu ile ile Glu 100 105 110

Cys Leu His val His val Phe Gin Leu val Gly ser Gly Phe Pro Asp 115 120 125

Asp ser pro val Met Thr Asn Ala Leu Thr Gly Leu Asp Arg ser Val 130 135 140

Ser Lvs Leu Met cys Thr Ser Asp Asp Lys Leu val Glu Ser val His 145 150 155 160

Tro ser Tyr Arg Thr ser ser Gly Gly Arg Tyr Arg Ala Thr val Gin 165 170 175

Thr Asn Phe Thr Phe Ala Lys Pro ile Glu Ala Gly Leu Lys Asn Asn 180 185 190

Met Pro Met Phe val Phe Arg Gin Leu Glu val Thr Gly ser Lys Thr 195 200 205

Glu Ile Gly Leu Gin Glu Gin Gin Lys Ala Phe Ser Thr Val Leu Met 210 215 220

Arg Leu Trp Leu Lys Cys Lys Arg Val Glu Ile Leu 225 230 235

<210> 30 <211 >243 <212> PRT <213> Branchiostoma floridae <400> 30 ser val pro Thr Asn Leu Asp Leu His ile Tyr Gly ser ile Asn Gly 1 5 10 15

Met Glu Phe Asp Met val Gly Gly Gly ser Gly Asn Pro Lys Asp Gly 20 25 30 ser Leu Ala Val Asn Val Lys ser Thr Lys Gly Ala Leu Arg Val ser 35 40 45

Pro Leu Leu val Gly pro His Leu Gly Tyr Gly His Tyr Gin Tyr Leu 50 55 60 pro Phe Pro Asp Gly Ala Ser pro Phe Gin Ala Ala Val Asn Asn Gly 65 70 75 80

Gly Tyr Gin Met His Arg ser Phe Asn Phe Glu Asp Gly Ala val Leu 85 90 95

Thr Ala Thr Tyr Asn Tyr Ser Tyr Ser Gly Gly Lys ile Gin Gly Glu 100 105 110

Phe His Lys Glu ile ile Glu Cys Leu His val His Val Phe Gin Leu 115 120 125

Val Gly ser Gly phe Pro Asp Asp ser Pro val Met Thr Asn Ala Leu 130 135 140

Thr Gly Leu Asp Arg ser Val Ser Lys Leu Met cys Thr ser Asp Asp 145 150 155 160

Lys Leu Val Glu ser val His Trp ser Tyr Arg Thr Ser Ser Gly Gly 165 170 175

Arg Tyr Arg Ala Thr val Gin Thr Asn Phe Thr phe Ala Lys Pro ile 180 185 190

Ala Ala Gly Leu Lys Asn Asn Met Pro Met Phe val Phe Arg Gin Leu 195 200 205

Glu Val Thr Gly ser Lys Thr Glu Ile Gly Leu Gin clu Gin Gin Lys 210 215 220

Ala Phe ser Thr Val Leu Met Arg Leu Trp Leu Lys cys Lys Arg Val 225 230 235 240

Glu ile Leu

<210> 31 <211 >243 <212> PRT <213> Branchiostoma floridae <400> 31

Met Ser val pro Thr Asn Leu Asp Leu His ile Tyr Gly ser ile Asn 15 10 15

Gly Met Glu Phe Asp Met Val Gly Gly Gly ser Gly Asn pro Asn Asp 20 25 30

Gly ser Leu ser val Asn val Lys Ser Thr Lys Gly Ala Leu Arg val 35 40 45

Ser Pro Leu Leu val Gly Pro His Leu Gly Tyr Gly His Tyr Gin Tyr 50 55 60 ueu Pro phe Pro Asp Gly Pro ser Pro phe Gin Ala Ala Val Asn Asn 65 70 75 80

Gly Gly Tyr Gin Met His Arg ser phe Asn Phe Glu Asp Gly Ala val 85 90 95 L.eu Thr Ala Thr Tyr Asn Tyr ser Tyr ser Gly Gly Lys ile Gin Gly 100 105 110

Glu Phe His val val Thr Glu cys Leu His Val His Val Phe Gin Leu 115 120 125 val Gly ser cys Phe Pro Asp Asp Ser pro Val Met Thr Asn Ala Leu 130 135 140

Thr Gly Leu Asp Arg ser Val Ala Lys Leu Met Cys Val Ser Asp Asp 145 150 155 160 ivs Leu Ala Glu phe val Asp Trp Thr Tyr Arg Thr ser Ser Gly Gly 1 165 170 175

Arg Tyr Arg Ala Thr val Gin Thr Asn Phe Thr Phe Ala Lys Pro ile 180 185 190

Ala Ala Gly Leu Lys Asn Asn Met pro Met Phe val Phe Arg Gin Leu 195 200 205

Glu Val Thr Gly ser Lys Thr Glu ile Ser Leu Gin Glu Gin Gin Lys 210 215 220

Ala Phe ser Thr val Leu val Arg Leu Trp Leu Lys cys Lys Arg Ala 225 230 235 240

Glu ile Leu

<210> 32 <211 >220 <212> PRT <213> Branchiostoma floridae <400> 32

Met pro Leu Pro Lys Thr His Glu Leu His ile Phe cly ser Phe Asn 1 5 10 15

Gly val Glu Phe Asp Met val Gly Arg cly ile sly Asn Pro Asn Glu 20 25 30

Gin Gly Ser Glu Glu Leu Asn Ala Lys phe Thr Lys Gly Pro Leu Lys 3 3 *rU ^ ^

Phe ser Pro Tyr ile Leu val Pro His Leu Gly grr Ala Tyr Tyr Gin

Tyr Leu Pro Phe Pro Asp Gly Met Ser Pro phe Gin Ala Ala Met His 65 70 75 80

Asp Gly ser Gly Tyr Gin Val His Arg Thr. ile Gin Tyr Glu Asp Gly 85 90 95

Ala Ser val Thr Ala His Tyr Arg Tyr Thr Tyr Glu Gly ser His ile 100 105 7 3 no

Lys Gly Glu phe Gin val ile Gly Thr Gly Phe Pro Pro Asp Gly Pro 115 120 125 val Met Thr Asn Lys Leu Thr Ala Met Asp Trp ser val Thr Lys Met 130 135 140

Leu Tyr Pro Asn Asp Lys Thr ile Leu ser Thr val Asp cys ser Tyr 145 150 155 160

Thr Thr Thr Glu Gly Lys Arg Tyr Gin ser Lys Met Arg Glu Asn Asn 165 170 175

Thr Phe Ala Lys Pro Met Ala Ala Asp ile Leu Gin Lys Gin Pro Met 180 185 190

Phe Val phe Arg Lys Thr Glu Leu Gin His ser Lys Thr Glu Leu Thr 195 200 205

Phe Lys Glu Trp Gin Lys Ala phe Thr Asp val Met 210 215 220

<210> 33 <211 > 231 <212> PRT <213> Branchiostoma floridae <400> 33

Met Pro Leu pro Lys Thr His Glu Leu His lie Phe Gly Ser Phe Asn 15 10 15

Gly val Lys phe Asp Met val Gly Glu Gly Thr Gly Asn Pro Asn Glu 20 25 30

Gly ser Glu Glu Leu Lys Leu Lys ser Thr Asn Gly pro Leu Lys Phe 35 40 45

Ser Pro Tyr lie Leu Val Pro His Leu Gly Tyr Ala Phe Asn Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Met Ser Pro Phe Gin Ala Ala Met Gin Asp 65 70 75 80

Glu ser Gly Tyr Gin val His Arg Thr Leu Gin Tyr Glu Asp Gly Ala 85 90 95

Phe val Thr Ala Asn Leu Arg Tyr Thr Tyr Glu Gly ser His lie Lys 100 105 110

Gly Glu Phe Gin His Tyr Asn Val Asp Tyr Asn lie lie Leu Gin val 115 120 125 lie Gly Thr Gly Phe Pro Pro Asp Gly Pro val Met Thr Asn Lys Leu 130 135 140

Thr Ala Leu Asp Trp ser Val Val Lys Phe Val Tyr Pro Asn Asp Lys 145 150 155 160

Thr ile Leu ser Thr Phe Asp Lys Thr Tyr Thr Thr Thr Glu Gly Lys 165 170 175

Arg Tyr Gin cys Thr Phe Arg Glu Asn ser Thr Phe Ala Lys Pro Met 180 185 190

Ala Ala Asp lie Leu Gin Lys Gin Pro Met Phe lie Phe His Lys Thr 195 200 205

Giu Leu Gin His Ser Asn Asn Ala Glu Leu Thr Phe Lys Glu Lys Gin 210 215 220

Thr Ala Phe ser Asp Met Lys 225 ¥ 230

<210> 34 <211 >230 <212> PRT <213> Ranchiostoma floridae <400> 34

Pro Leu Pro Thr Thr His Glu Val His Val Tyr Gly ser He Asn Gly 15 10 15 val Glu Phe Asp Leu Val Gly ser Gly Lys Gly Astl Pro Lys Asp G-|n 20 25 30

Gly ser Glu Glu lie Gin Val Lys ser Thr Lys Gly Pro Leu Gly Phe 35 40 45 ser Pro Pro Val Val val Pro Asn lie Gly Tyr Gly Phe His Gin Tyr 5° 55 60

Leu Pro Phe Pro Asp Gly Met ser Pro Phe Gin Ala Ala Ala Asp Asp 65 70 75 μ 80

Gly Ser Gly Tyr val Val His Arg Asn lie·Gin Phe Glu Asp Gly Ala 85 90 95

Ser Leu Thr Gly lie Tyr Arg Tyr Ser Tyr Asp Ala Gly His He Lys 100 105 110

Gly Glu Phe Arg Tyr Val ser Asp lie phe Gin Val val Gly ser Gly 115 120 125

Phe pro Ala Asp Gly Pro val Met Thr Lys Ser Leu Thr Ala val asp 130 135 140

Trp ser Val Ala Thr Met Leu Phe pro Asn Asp Thr Thr Val val ser 145 150 155 160

Thr lie Asp Trp Thr cys Pro Thr Thr Ser Gly Lys Arg Tyr His Ala 165 170 175

Thr Val Arg Thr Asn Tyr Thr Phe Ala Lys Pro lie Ala Gly ser lie 180 185 190

Leu Gin Lys Gin Pro Met Phe val phe Arg Lys Thr Glu Val Lys Ala 195 200 205 ser Asp ser Glu lie Asn Leu Lys Glu ser Gin Lys Ala Phe His Asp 210 215 220

Leu val Gly lie cys lie 225 230

<210> 35 <211 > 218 <212> PRT <213> Branchiostoma floridae <400> 35

Met Pro Leu Pro Ala Thr His Glu lie His Leu His Gly ser val Asn 15 10 15

Gly His Glu Phe Asp Leu val Gly ser Gly Lys Gly Asp Pro Lys Ala

Gly ser Leu val Thr Glu Val Lys ser Thr Met Gly Arg Leu Lys Phe 35 40 45 ser Pro His Leu Met He Pro His Leu Gly Tyr Gly Tyr Tyr Gin Tyr 50 55 60

Leu Pro Tyr Pro Asp Gly Pro Ser Pro Phe Gin Thr Ala Met Leu Asp 65 70 75 80

Gly ser Gly Tyr Lys val His Arg Val Phe Asn Phe Glu Asp Gly Gly 85 90 95 val Leu ser lie Asp Tyr Asn Tyr Ala Tyr Glu Gly Thr His lie Lys 100 105 110

Ser Asp Phe Lys Leu Met Gly Ser Gly Phe Pro Asp Asp Gly Pro Val 115 120 125

Met Thr Ser Gin lie val Asp Gin Asp Gly cys val ser Lys Lys Thr 130 135 140

Tyr Leu Asn Asp Asn Thr Tie Val Asp ser Phe Asp Trp Ser Tyr Asn W5 150 155 160 leu Gin Asn Gly Lys Arg Tyr Arg Ala Arg val Thr ser Asn Tyr lie 165 170 175

Phe Gly Lys Pro Leu Ala Ala Asp Val Met Lys Lys Gin Pro Val Phe 180 185 190

Val Tyr Arg Lys cys Tyr val Lys ser Thr Gin Thr Glu lie Thr Leu 195 200 205

Asp Glu Arg Glu Lys Ala Phe Tyr Glu Val 210 215

<210> 36 <211 > 231 <212> PRT <213> Branchiostoma floridae <400> 36

Met ser val Pro Thr Asn Leu Asp Leu His lie Tyr Gly ser lie Asn 1 5 10 15

Gly Met Glu Phe Asp Met val Gly Gly Gly ser Gly ash Pro Lys Asp 20 25 30

Gly ser Leu ser val Asn val Lys ser Thr Lys Gly Ala Leu Arg Val 35 40 45 ser Pro Leu Leu val Gly Pro His Leu Gly Tyr Gly His Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Asp Pro ser pro Phe Gin Ala Ala val Asn Asn 65 70 75 80

Gly Gly Asn Gin Met His Arg ser Phe Asn Phe Glu Asp Gly Ala val 85 90 μ 95

Leu Thr Ala Thr Tyr Asn Tyr Ser Tyr ser Gly Gly Lys lie Gin Gly 100 105 110

Glu Phe His Leu val Gly Ser Cys Phe Pro Asn Asp ser Pro val Met 115 120 125

Thr Asn Ala Leu Thr Gly Leu Asp Arg ser val Ala Lys Leu Met cys 130 135 140

Val ser Asp Asp Lys Leu Ala Glu Phe val Asp Trp Thr Tyr Arg Thr 145 150 155 160

Ser ser Gly Gly Arg Tyr Arg Ala Thr val Gin Thr Asn Phe Thr Phe 165 170 175

Ala Lys Pro lie Ala Ala Gly Leu Lys Asn Asn Met Pro Met Phe Val 180 185 190

Phe Arg Gin Leu Glu val Thr Gly ser Lys Thr Glu lie Gly Leu Gin 195 200 205

Glu Gin Gin Lys Ala Phe Ser Thr Val Leu val Arg Ser Trp Leu Lys 210 215 220

Tyr Lys Arg Ala Glu lie Leu 225 230

<210> 37 <211> 218 <212> PRT <213> Branchiostoma floridae <400> 37

Met ser Leu Pro Thr Thr His Asp Leu His lie Phe Gly Ser val Asn 15 10 15

Gly Ala Glu Phe Asp Leu val Gly Gly Gly Lys Gly Asn ppo Asn ASP 20 25

Gly Thr Leu Glu Thr ser Val Lys Ser Thr Arg Gly A^a Leu Pro cys 35 40 45 ser Pro Leu Leu lie Gly Pro Asn Leu Gly Tyr Gly p^e Tyr Gln Tyr 50 55 60

Leu Pro Phe Pro Gly Gly Ala Ser Pro Phe Gin Τ^Γ A^a ^ T^r 65 70 75 8U

Gly Gly Tyr Gin val His Arg Val Phe Lys Phe Glu Asp Gly Gly val 85 90 95

Leu ser cys Asn Phe Arg Tyr Thr Tyr Glu Gly Gly Lys ile Lys Gly 100 105 uo

Glu Phe Gin Leu ile Gly ser Gly Phe Pro Ala Gly Gly Pro val Met 115 120 12¾ ser Gly Gly Leu Thr Thr Leu Asp Arg ser Val Ala Lys Leu Gin cys 130 135 140 ser Asp Asp Arg Thr ile Thr Gly Thr Asn Asn Trp ser Phe Trp Thr 145 150 155 H 160

Thr Asp Gly Lys Arg His Gin Ala Asp val Gin Thr Asn Tyr Thr Phe 165 170 r 175

Ala Lys pro Leu Pro Ala Gly Leu Lys Glu Lys Met Pro Ile Phe Leu 180 185 190

Gly His Gin ile Glu Val Lys Ala ser Lys Thr Glu ile Thr Leu ser 195 200 205

Glu Lys Val Lys Ala Phe ile Asp Thr Val 210 215

<210> 38 <211 >253 <212> PRT <213> Branchiostoma floridae <400> 38

Met His Ala Thr Lys His Thr Val Gly cys Tyr Pro Leu Tyr Leu Gly 15 10 15

His Gin ser Leu Pro Thr Thr His Asp Leu His ile phe Gly ser val 20 25 30

Asn Gly Ala Glu Phe Asp Leu Val Gly Gly Gly Lys Gly Asn Pro Asn 35 40 ij* 45*

Asp Gly Thr Leu Glu Thr ser val Lys ser Thr Arg Gly Ala Leu Pro 50 55 60

Cys ser Pro Leu Leu ile Gly pro Asn Leu Gly Tyr Gly Phe Tyr Gin 65 70 75 80

Tyr Leu Pro Phe Pro Gly Gly Ala Ser Pro Phe Gin Thr Ala ile Thr 85 90 95

Asp Gly Gly Tyr Gin Val His Arg Val phe Lys Phe Glu Asp Gly Gly 100 105 110

Val Leu Asn Cys Asn Phe Arg Asn Phe Arg Tyr Thr Tyr Glu Gly Gly 115 120 125

Lys ile Lys Gly Glu phe Gin Len Ile Gly ser Gly phe Pro Ala Gly 130 135 140

Gly Pro val Met Pro Gly Gly Leu Thr Thr Leu Asp Arg Ser val Ala 145 150 155 160

Lys Leu Gin cys ser Asp Asp Arg Thr ile Thr Gly Thr Asn Asn Trp 165 170 175 ser Phe Trp Thr Thr Asp Gly Lys Arg His Gin Ala Asp val Gin Thr 180 185 190

Asn Tyr Thr ser Pro Ser Arg ser Gly Arg Ser Gin Gly Glu Asp Ala 195 200 205

Gly Leu Pro Gly Ala Pro Asp Arg Gly Gin Ser Val Gin Asp Arg Asp 210 215 220

His Pro Glu Arg Glu ser Glu Gly Leu His Arg His cys val Lys Phe 225 230 235 240

Lys Phe Ala Asp cys Val Lys Pro Arg ile Gin ser cys 245 250

<210> 39 <211> 221 <212> PRT <213> Branchiostoma floridae <400> 39 ser Leu Pro Thr Ala His Asp Leu His ile Phe Gly ser val Asn Gly 1 5 10 15

Ala Glu Phe Asp Leu Val Gly Gly Gly Lys Gly Asn Pro Asn Asp Gly 20 25 30

Thr Leu Glu Thr Ser Val Lys Ser Thr Arg Gly Ala Leu Pro Cys Ser 35 40 45

Pro Leu Leu Ile Gly pro Asn Leu Gly Tyr Gly Phe Tyr Gin Tyr Leu 50 55 60 pro phe Pro Gly Gly Ala ser Pro Phe Gin Thr Ala ile Thr Asp Gly 65 70 75 80

Gly Tyr Gin val His Arg Val Phe Lys Phe Glu Asp Gly Gly val Leu 85 90 95

Asn cys Asn Phe Arg Tyr Thr Tyr Glu Gly Gly Lys ile Lys Gly Glu 100 105 110

Phe Gin Leu Ile Gly ser dly phe Pro Ala Gly Gly pro val Met ser 115 120 125

Gly Gly Leu Thr Thr Leu Asp Ara ser val Ala Lys Leu Gin cys Ser 130 135 140

Asp Asp cys Thr ile Thr Gly Thr Asn Asn Trp ser Phe cys Thr Thr 145 150 155 160

Asp Giv Lys Arg His Gin Ala Asp Val Gin Thr Asn Tyr ile Phe Ala 165 170 175

Lys pro Leu Pro Ala Gly Leu Lys Glu Lys Met Pro ile phe Leu Giv 180 185 190

His Gin Ile Glu val Lys Ala ser Lys Thr Glu ile Thr Leu cys Glu 195 200 205

Lys val Lys Ala Phe ile Asp Thr Asp Leu Leu Glu Ile 210 215 220

<210> 40 <211 >222 <212> PRT <213> Branchiostoma floridae <400> 40

Met ser Pro ile ser Pro Leu Pro Thr Thr His Glu Val His val Tyr 1 5 10 15

Gly Ser ile Asn Gly Val Glu Phe Asp Leu Val Gly ser Giv lvs Gly 20 25 30

Asn Pro Lys Asp Gly Ser Glu Glu ile Gin val Lys ser Thr lvs Gly 35 40 45

Pro Leu Gly Phe Ser Pro Tyr Ile Val val pro Asn Ile Gly Tyr Gly 50 55 60

Phe His Gin Tyr Leu Pro Phe Pro Asp Gly Met Ser Pro phe- Gin Ala 65 70 75 80

Ala Ala Asp Asp Gly ser Gly Tyr val val His Arg Thr ile Gin Phe 85 90 95

Glu Asp Gly Ala ser Leu Thr Gly Asn Tyr Arg Tyr ser Tyr Asp Gly 100 105 110

Gly His Ile Lys Gly Glu Phe His Val val Gly ser Gly Phe Leu Gly 115 120 125

Asp Gly Pro val Met Thr Lys ser Leu Thr Ala val Asp Trp ser val 130 135 140

Ala Thr Met Leu Phe Pro Asn Asp Thr Thr Val val ser Thr ile Asp 145 150 155 160

Trp Thr cys Pro Thr Thr ser Gly Lys Arg Tyr His Ala Thr Val Arg 165 170 175

Thr Asn Tyr Thr Phe Ala Lys Pro Ile Ala Ala ser ile Leu Gin Lys 180 185 . 190

Gin Pro Met Phe val Phe Arg Lys Thr Glu Val Lys Ala ser Asp Ala 195 200 205

Glu ile Asn Leu Lys Glu Trp Gin Lys Ala Phe His Asp Leu 210 215 220

<210> 41 <211 >220 <212> PRT <213> Branchiostoma floridae <400> 41

Met Pro Leu Pro Thr Thr His Glu Leu His ile phe Giv ser Phe Asn 1 5 10 J 15

Gly val Glu phe Asp Met val Gly Arg Gly He cly Asn pro Asn Asp 20 25 30

Gly Tyr Glu Glu Leu Asn Leu Lys ser Thr Lys Gly Ala Leu Lys Phe 35 40 45 ser Pro Trp lie Leu val Pro Gin lie Gly Tyr Gly Phe His Gin Tvr 50 55 60

Leu Pro Tyr Pro Asp Gly Met ser Pro Phe Gin Ala Ala Met Gin Asp 65 70 75 80

Gly ser Gly Tyr Gin Val His Arg Thr Met Gin Phe Glu Asp Gly Ala 85 90 95

Ser Leu Thr Ala His Phe Arg Tyr Thr Tyr Glu Gly ser His lie Lys 100 105 no

Gly Glu Phe Gin val lie Gly Thr Gly Phe Pro Ala Asp Glv Pro val 115 120 125

Met Thr Asn Lys Leu Thr Ala Ala Asp Trp Cys val val Lys Met Val 130 135 140

Tyr Pro Asn Asp Lys Thr lie Leu ser Thr Phe Asp Trp Thr Tvr Thr 145 150 155 160

Thr Glu Gly Lys Arg Tyr Gin Ser Thr Val Arg Thr Asn Tyr Thr Phe 165 170 175

Ala Lys pro Met Ala Ala Asn lie Leu Gin Lys Gin pro Met phe val 180 185 190 phe Arg Lys Thr Glu Leu Gin His ser Lys Thr Glu Leu Thr Phe Lys 195 200 205

Glu Trp Gin Lys Ala Phe Thr Asp val Met val Phe 210 215 220

<210> 42 <211> 169 <212> PRT <213> Branchiostoma floridae <400> 42

Met Pro Leu Pro Ala Thr His Glu Ile His ile Tyr Gly ser Val Asn 1 5 10 15

Gly His Glu phe Asp Leu Val Gly Gly Gly Lys Gly Asp pro Asn Ala 20 25 30

Gly Ser Leu val Thr Glu val Lys Ser Thr Met Gly Pro Leu Lys Phe 35 40 45 ser Pro His Leu Met lie pro His Leu Gly Tyr Gly Tyr Tyr Gin Tyr 50 55 60

Leu Pro Tyr Pro Asp Gly Pro Ser pro Phe Gin Thr Ala Met Leu Asp 65 70 75 80

Gly Ser Gly Tyr Ser Val His Arg Val Phe Asp Phe Glu Asp Gly Gly 85 90 95

Lys Leu Thr Leu Glu Phe Lys Tyr Ser Tyr Glu Gly Ser His lie Lys 100 105 110

Ala Asp Met Lys Phe Thr Gly Ser Gly phe Pro Asp Asp Gly Pro val 115 120 125

Met Thr ser Gin lie val Asp Glu Asp Gly cys val ser Lys Asn Thr 130 135 140 lie His Asn Asp Asn Thr lie val Asp Asn Phe Asp Trp Thr Asn val 145 150 155 160

Leu Gin Asn Gly Lys Arg Tyr Arg Ala 165

<210> 43 <211 >233 <212> PRT <213> Branchiostoma floridae <400> 43

Met Pro Leu Pro Ala Thr His Glu He His Leu His Gly ser lie Asn 1. 5 10 15

Gly His Glu Phe Asp Leu Ala Gly Gly Gly Lys Gly Asp Pro Asn Ala 20 25 30

Gly ser Leu val Thr Thr Ala Lys ser Thr Lys Gly pro Leu Lvs Phe 35 40 45 y ser pro His Leu Met lie Pro His Leu Gly Tyr Gly Tyr Tyr Gin Tyr

Leu Pro Tyr Pro Asp Gly Pro ser Pro Phe Gin Ala Thr Met Leu Glu 65 70 75 80

Gly ser Gly Tyr Thr Val His Arg val Phe Asp Phe Glu Asp Gly Gly

Lys Leu Ser ile Glu Phe Lys Tyr ser Tyr Glu Gly ser His ile lvs 100 105 110

Ala Asp Met Lys Arg ser Thr ser Asp Thr Asn val His val Phe Leu 115 120 125

Phe Gin Phe Thr Gly Thr Gly Phe pro Glu Asp Gly pro Val Met Thr 130 135 140 ser Gin Ile Val Asp Gin Asp Gly cys Val ser Lys Asn Thr Tvr Leu 145 150 155 * 160

Asn Asp Asn Thr ile Val Asp Asn Phe Asp Trp Thr Tyr Asn Leu Gin 165 170 175

Asn Gly Lys Arg Tyr Arg Ala Arg val Thr ser His Tyr ile Phe asd 180 185 190

Lys Pro Phe ser Ala Asp Leu Met Lys Lys Gin Pro val Phe val Tvr 195 200 205

Arg Lys cys His val Lys Ala ser Lys Thr Glu ile Asn Leu Asp Glu 210 215 220

Arg Glu Lys Ala Phe Tyr Glu ser Ala 225 230

<210> 44 <211> 219 <212> PRT <213> Branchiostoma floridae <400> 44

Met Pro Leu Pro Ala Thr His Asp ile His Leu His Gly Ser ile Asn 15 10 15

Gly His Glu Phe Asp Met val Gly Gly Gly Lys Gly Asp Pro Asn Ala 20 25 30

Gly ser Leu Val Thr Thr Ala Lys Ser Thr Lys Gly Ala Leu Lys Phe 35 40 45

Ser Pro Tyr Leu Met Ile Pro His Leu Gly Tyr Gly Tyr Tyr Gin Tyr 50 55 60

Leu Pro Tyr Pro Asp Gly Pro ser pro phe Gin Thr ser Met Leu Glu 65 70 75 80

Gly ser Gly Tyr Ala val Tyr Arg Val Phe Asp Phe Glu Asp Gly Gly 85 90 95

Lys Leu Thr Thr Glu Phe Lys Tyr Ser Tyr Glu Gly ser His ile Lys 100 105 110

Ala Asp Met Lys Leu Met Gly Ser Gly Phe pro Asp Asp Gly Pro Val 115 120 125

Met Thr ser Gin ile val Asp Gin Asp Gly Cys Val Ser Lys Lys Thr 130 135 140

Tyr Leu Asn Asn Asn Thr Ile val Asp ser Phe Asp Trp Ser Tyr Lys 145 150 155 160

Leu Gin Asn Gly Lys Arg Tyr Arg Ala Arg val ser Ser His Tyr lie 165 170 1/5

Phe Asp Lys Pro Phe ser Ala Asp Leu Met Lys Lys Gin pro val Phe H 180 185 190

Val Tyr Arg Lys cys His val Lys Ala ser Lys Thr Glu val Thr Leu 195 200 205

Asp Glu Arg Glu Lys Ala Phe Tyr Glu Leu Ala 210 215

<210> 45 <211> 211 <212> PRT <213> Branchiostoma floridae <400> 45

Met Pro Leu Pro Ala Thr His Glu lie His Leu His Gly Ser Val Asn 15 10 15

Gly His Glu Phe Asp Leu val Gly Gly Gly Lys Gly Asp Pro Lys Ala 20 25 30 ser Thr Lys Gly Pro Leu Lys Phe ser Pro His Leu Met lie Pro His 35 40 45

Leu Gly Tyr Gly Tyr Tyr Gin Tyr Leu Pro Tyr Pro Asp Gly Pro ser 50 55 60

Pro Phe Gin Thr Ala Met Leu Asp Gly ser Gly Tvr Lvs Val His Arq 65 70 75 80 val Phe Asn Phe Glu Asp Gly Gly Val Leu ser lie Glu Tyr Asn Tyr 85 90 95 ser Tyr Glu Gly Thr His lie Lys ser Asp Phe Lys Leu Met Gly ser 100 105 110

Gly Phe Pro Asp Asp Gly Pro Val Met Thr ser Gin lie Val Asp Gin 115 120 125

Asp Gly cys val ser Lys Lys Thr Tyr Leu Asn Asp Asn Thr lie val 130 135 145

Asp ser Phe Asp Trp ser Tyr Lys Leu Gin Asn Gly Lys Arg Tyr Arg 145 150 155 160

Ala Arg Val Thr ser Asn Tyr He Phe Gly Lys Pro Leu Ala Ala Asp 165 170 175 val Met Lys Lys Gin Pro val Phe val Tyr Arg Lys cys Tyr val Lys 180 185 190

Ser Thr L^s Thr Glu lie Thr Leu Asp Glu Arg Glu l^s Ala Phe Tyr

Glu Leu Ala 210

<210> 46 <211 > 218 <212> PRT <213> Branchiostoma floridae <400> 46

Pro Leu pro Thr Thr His du Leu His ile Phe Gly Thr Phe Asn Gly 15 10 15 val Glu Tyr Asp Met val Gly Arg Gly Lys dy Asn Pro Asn Asp Gly 20 25 30

Tyr Glu Glu Leu Asn Leu Lys ser Thr Lys Gly Pro Leu Lys Phe ser 35 40 45

Pro Trp ile Leu val Pro Gin Ile Gly Tyr Gly Phe His Gin Tyr Leu 50 55 60 pro Tyr pro Asp Gly Met Ser Pro phe Gin Ala Ala Met His Asp Gly 65 70 75 80 ser Gly Tyr Gin Val His Arg Thr Leu Asp Phe Glu Asp Gly Ala Thr 85 90 95

Leu Thr Ala Asp phe Arg Tyr Thr Tyr Glu Gly ser His Ile Lys Gly 100 105 110

Glu Phe Lys Val Ile Gly Thr Gly Phe Pro Ala Asp Gly Pro Val Met 115 120 125

Thr Asn Lys Leu Thr Ala Ala Asp Trp cys Val Asn Lys Met Leu Tyr 130 135 140

Pro Asp Asp Lys Thr Ile Asn Ser Thr Phe Asp Trp Ser Tyr Thr Thr 145 150 155 160

Ser Glu Gly Lys Arg Tyr Gin ser Thr Val Arg Glu Asn Tyr Thr Phe 165 170 175

Ala Lys Pro Met Ala Ala Asn ile Leu Gin Lys Gin Pro Met Phe Val 180 185 190

Phe Arg Lys Thr Glu Leu Gin His ser Lys Thr Glu Leu Thr Phe Lys 195 200 205

Glu Trp Gin Lys Ala Phe Thr Asp Val Met 210 215

<210> 47 <211 > 11 <212> PRT <213> Branchiostoma sp. <220> <221 > VARIANT <222> (3)..(10) <223> Xaa in position 3 is Ala or Gly Xaa in position 4 is Phe, His or Tyr Xaa in position 5 is His, Tyr or Asn Xaa in position 10 is Phe or Tyr <400> 47

Gly Tyr Xaa xaa xaa Gin Tyr Leu Pro xaa Pra 15 10

<210> 48 <211> 1570 <212> DNA <213> Branchiostoma lanceolatum <400> 48 acaatcatgc ctctcccagc aacccacgag ttacacatct ttggctccat caatagtttg 60 gagtrtgacc tggtgggtcg tggcaccggc aacccaaagg aaggttatga ggaactccac 120 ccgaagtcca ccaagagtgc cctccagttc tccccatgga tcctggtccc tcaaatcggg 180 tacggctttt accagtacct gcccttcccc gatggagcga tgtcgccttt tcaggccgct 240 atgaaegatg gctccggata ccaagtccat cgcacaatgc agtttgaaga cggtgcaacc 300 ctgactggca tctaccgcta tacctacgag ggaacccaca tcaaaggaga gtttcaggtg 360 atcgggactg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgcgtga ccaagattgt atacccgaac gagaatacca tcatcgacaa attcgactgg 480 acctacacca ctacaagtgg caagcgctac cagagcaatg tgcggtccaa cttcaccttt 540 gccaagccga tcgcggccaa catcctgcag aagcagccga tgttcgtgtt ccgtaagacg 600 gagctaaagc actccaagac cgagctcaac ttcaaggagt ggcagacggc ctttagcgat 660 gtgatgtgag catctagtgt attttcacat ttggctggga aatacccaaa gaaacatgtc 720 cattttcttt .atagaaccca attctgatat ggagcaagga cataaaacat tttcacctac 780 gagtattatt cgtctgtcaa ttttcattgt atttgtttga aaacttatat tatcgaacta 840 tgttttaacc attggactac agacttttgg caggcatgct ttagagcctt tattttagaa 900 tgacacttta ctcaacgacg aaggtactaa tactacttcc tctggtcatt gtttttactt 960 i ggaaagggtg tttaaacagt caaatgtaaa taagattgga tgtttttcat catagttgtt 1020 ttgatactgc ggtgaatgcg tgctttcgct atcttatata ttacaataca atacgaccac 1080 atatgattag cagaacagta ctacgagttt acctttagga tcaaaatgtc agattacgtt 1140 aattctttcc taatcaagtg gatgtagtag aggttgtacc gccttagcag aagacaaaat 1200 gaaaacataa aaatacaaat acttgacgga cgtgcagtca ttctctcatt ggtcgaaccg 1260 ataaatgtga tggacagtca ggatcagtct attacggctt ggattttcta tctgttctca 1320 aaacacaaag acatcgtatc tgtgctcctt taacgtcgat gtgtagtggt attgtgttat 1380 taaagtttat tgtgtaggaa ttactagaaa ttggagtatt ttctagtcaa gcctctagcc 1440 tcataaaatg ctctggatgc cataaaagat ctatctgttt gctgccatta ttgtagttat 1500 tttgccttct tgttattgca aataaaggct atgggtgcta ctgcttaaaa aaaaaaaaaa 1560 aaaaaaaaaa 1570 <210> 49 <211 > 1120

<212> DNA <213> Branchiostoma lanceolatum <400> 49 acaaccatgt ctctcccagc gacccacgag ttacacatct ttggctccat caatagtttg 60 gagtttgacc tggtgggtcg tggcaccggc aacccaaggg aaggttatga ggaactccac 120 ctgaagtcca ccaagagtgc cctccagttc tccccatgga tcctggtccc tcaaatcggg 180 tacggctttt accagtacct gcccttcccc gatggagcga tgtcgccttt tcaggccgct 240 atgaacgatg gctccggata ccaagtccat cgcacaatgc agtttgaaca cggtgcaacc 300 ctgactggca tctaccgcta tacctacgag ggaacccaca tcaaaggaga gtttcaggtg 360 atcgggactg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgcgtga ccaagattgt atacccgaac gagaatacca tcatcgacaa attcgactgg 480 acctacacca ctacaagtgg caagcgctac catagcaatg tgcggtccaa cttcaccttt 540 gccaagccga tcgcggccaa catcctgcaa aagcagccga tgttcgtgtt ccgtaagacg 600 gagctaaagc attccaagac cgagctcaac ttcaaggagt ggcagacggc ctttggcgat 660 gtgatgtgag catctactgt attttcccat ttggctggga aataccacac aaacatgtcc 720 attttcttta tagaacccaa ttctgatatg gagcaaggga cataacaaca ttttcaccta 780 ccgaataata ttcgtctgtc aattctcatt gtaacttgtt tgaaaactta tattatcgaa 840 ctatgttctt acccattgga actaccgacc ttttggcagg catgcttata gatcctttta 900 tttttagaat gacactctac tcaacgacga aggcactaat acctacttcc tcagtccatt 960 gttattacct gggaaatgtg tttaaacagt ccaaatgcca aataagaatt gaatggtctt 1020 gcaacatact tgcttttgaa cacttgccag ctaaatgcct gcctatcgcc tatcgtatta 1080 tttctacaag taaaagatta cggacccacc tatatggatt 1120

<210> 50 <211 >775 <212> DNA <213> Branchiostoma lanceolatum <400> 50 acaaccatgt ctctcccagc gacacatgag ttacacatct ttggctcctt caacggtgtg 60 gactttgaca tggtgggtcg tggcaccggc aatccaaatg atggttatga ggagttaaac 120 ctgaagtcca ccaagggtgc cctccagttc tccccctgga tcctggtccc tcaaatcggg 180 tatggcttcc atcagtacct gcccttcccc gacgggatgt cgcctttcca ggccgccatg 240 aaagatggct ccggatacca agtccatcgc acaatgcagt ttgaagacgg tgcctccctg B00 acttccaact accgctacac ctacgaggga agccacatca aaggagagtt tcaggtgatc 360 gggactggtt tccctgctga cggtcctgtg atgaccaact cgctgaccgc tgcggactgg 420 tgcgtgacca agatgctgta ccccaacgac aaaaccatca tcagcacctt tgactggact 480 tacaccactg gaagtggcaa gcgctaccag agcacagtgc ggaccaacta cacctttgcc 540 aagccaatgg cggccaacat cctgaagaac cagccgatgt tcgtgttccg taagacggag 600 ctcaagcact ccaagaccga gctcaacttc aaggagtggc aaaaggcctt taccgatgtg 660 atgtgagcgt ecagtttgtt acacttgact gggcccacac caaaagaaat attactcatt 720 agtttctgat aaaacccaaa aactgcagca aagagataaa acatttgtca cctac 775

<210> 51 <211 >893 <212> DNA <213> Branchiostoma lanceolatum <400> 51 acagtcatgc ctctcccagc aacacatgag ttacacatct ttggctcctt caacggtgtg 60 gactttgaca tggtgggtcg tggcaccggc aatccaaatg atggttatga ggagttaaac 120 ctgaagtcca ccaagggtgc cctccagttc tccccctgga tcctggtccc gcaaatcggg 180 tatggcttcc atcagtatct gcccttcccc gacgggatgt cacctttcca ggctgccatg 240 aaagatggct ccggatacca agtccatcgc acaatgcagt ttgaagacgg tgcctccctg 300 acttccaact accgctacac ctacgaggga agccacatca aaggagagtt tcaggtgatc 360 gggactggtt tccctgctga cggtcctgtg atgaccaact cgctgaccac tgcggactgg 420 tgcgtgacca agatgctgta ccccaacgac aaaaccatca tcagcacctt tgactggacc 480 tacaacactg caagtggcaa gcgctaccag agcacggtgc gaaccaacta cacctttaac 540 aagccaatgg cggccaacat cctgaagaac cagccgatgt tcgtgttccg caagacggag 600 ctcaagcact ccaagaccga gctcaacttc aaggagtggc aaatggcctt tgccgatgtg 660 atgtgagcgt ccggtgtgtt acacttgact ggaaacaaac caaaagaaac attacacatt 720 agtttatggt aaaaccaaat actgcagcaa agatatacaa aattgtcacc tacgaaatgt 780 caatgttcat tgttaaaaaa aatttatgat atccaagata tttacaccat ttgggctgca 840 aactgttggc aggcatgctt tagggtcatt attttcagaa tgggcaccta acg 893 <210> 52 <211 > 1210

<212> DNA <213> Branchiostoma lanceolatum <400> 52 acagtcatgc ctctcccagc aacacacgag ttacacatct ttggttcctt caatggtgtg 60 gactttgaca tggtgggtca tggcaccggc aatccaaatg atggttatga ggagttaaac 120 ctgaagtcca ccaagggtgc cctccagttc tccccctgga tcctggtccc tcaaatcggg 180 tatggcttcc accagtacct gcccttcccc gacgggatgt cgccttttca ggctgccatg 240 aaagatggct caggatacca agtccatcgc acaatgcagt ttgaagacgg tgcctccctg B00 acttccaact accgctacac ctacgaggga agccacatca aaggagagtt tcaggtgaac 360 gggactggtt tccctgctga cggccctgtg atgaccaact cgctgaccac tgcggactgg 420 tgcgtgacca agatgctgta tcctaacgac aagaccatxa tcagcacctt tgactggacc 480 tacaccactg gaagtggcaa gcgctaccag agcacagtgc ggaccaacta cacctttgcc 540 aagccaatgg cggccaacat cctgcagaac cagccgatgt tcgtgttccg caagacggag 600 ctcaagcact ccaagaccga gctcaacttc aaggagtggc aaaaggcttt tgccgatgtg 660 atgtgagcgt ccagtttgtt acacttgact ggaaacaaac caaaagaaac attacacatt 720 agtttatggt aaaaccaaat actgcagcaa agatatacaa aattgtcacc tacgaatgtc 780 aatgttcatt gttaaaaaaa acttatgata tccaagatat ttacaccatt tggctgcaaa 840 ctgttgggca ggcatgcttt agggtcatta tttcagaatg gcacctacga aggtactaat 900 actgcttcct ctggtcattt ttttcacttc gaaagggcgt tttaacagtc aagtgtaaat 960 ataattggat gtttgtcatc atgattgctt tgatactgcg gtgaatgcgt gctttcgctt 1020 ttcttatatt ataacaatac taccttatgt gaataagcaa ggattgctag gtacggagct 1080 taccttagga tcggaaatgt tcagattacc tttaaatttt tcctgatcaa ttggatgtag 1140 tgaatggcct accagtccac cctagctccc atttacaaga tgactaaatt tttcttacac 1200 ccgggcggtc 1210

<210> 53 <211 > 891 <212> DNA <213> Branchiostoma lanceolatum <400> 53 acaaccatgt ctctcccagc gacccacgag ttacacatct ttggctccat caatagtttg 60 gagtttgacc tggtgggtcg tggcaccggc aacccaaagg aaggttatga ggaactccac 120 ctgaagtcca ccaagagtgc cctccagttc tccccatgga tcctggtccc tcaaatcggg 180 tacggctttt accagtacct gcccttcccc gatggagcga tgtcgccttt tcaggccgct 240 atgaacgatg gctccggata ccaagtccat cgcacgatgc agtttgaaga cggtgcaacc 300 ctgactggca tctaccgcta tacctacgag ggaacccaca tcaaaggaga gtttcaggtg 360 atcgggactg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgcgtga ccaagattgt atacccgaac gagaatacca tcatcgacaa attcgactgg 480 acctacacca ctacaagtgg caagcgctac cagagcaatg tgcggtccaa cttcaccttt 540 gccaagccga tcgcggccaa catcctgcag aagcagccga tgttcgtgtt ccgtaagacg 600 gagctaaagc actccaagac cgagctcaac ttcaaggagt ggcagacggc ctttagcgat 660 gtgatgtgag catctagtgt attt'tcacat ttggctggga aatacccaaa gaaacatgtc 720 cattttcttt atagaaccca attctgatat ggagcaagga cataaaacat tttcacctac 780 gagtattatt cgtctgtcaa ttttcattgt atttgtttga aaacttatat tatcgaacta 840 tgttttaacc attggactac agacttttgg caggcatgct ttagagccct c 891

<210> 54 <211 >882 <212> DNA <213> Branchiostoma lanceolatum <400> 54 acaaacatgc ctcttccagc gacccatgag ttacacatct ttggctcctt caatggtgtg 60 gagtttgaca tggttggtcg cggcactggc aacccaaatg atgggtctga ggatttacac 120 ctgaagtcca ccaagggtgc cctccagttc tccccctgga tcctaatccc tcacatcggg 180 tacggctttc acgagtacct gccctttccc gacgggatgt cgcctttcca ggccgccatg 240 caagacggct ccggatacca agtgcatcgc actatgcagt ttgaagacgg tgcctccctg 300 actgcaaact tccgctacac ctacgaggga agccacatca aaggagagtt tcaggtgatc 360 gggtccggtt tccctgctga cggccctgtg atgaccaact cgctgaccgc tgtggactgg 420 tgcgtggcca agatgctgta ccccaacgac aagaccatca taagcacctt tgactggacc 480 tacaccactg gaagtggcaa gcgctaccag agcacagtgc ggaccaacta cacctttgcc 540 aagccaatgg cggccaacat cctgaagaac cagccgatgt tcgtgttccg caagacggag 600 ctcaagcact ccaagaccga gctcaacttc aaggagtggc aaaaggcctt tgccgatgtg 660 atgtgagcgt ccggtgtgtt acacttgact ggaaacaaac caaaagaaac gttacacatt 720 agtttatggt aaaaccaaat actgcagcaa agatatacaa aattgtcacc tacgaatgtc 780 aatgttcatt gttaaaaaaa aaacttatga tatccaagat atttacacca tttggctgca 840 aactgttggc aggcatgctt tagggtcatt atttcagaat gg 882

<210> 55 <211> 1239 <212> DNA <213> Branchiostoma lanceolatum <400> 55 acaaacatgc ctcttccagc gacccacgat ttacacatct ccggctcaat caatggacat 60 gagtttgact tggaagggtc tggtaagggc aatgaaaaag aaggttatca ggagctccac 120 ctaaagtcca acaagggtga cctgtcattc tccccctgga tcctggtccc aaacaccggc 180 tacggtttct accagtacct gcccttcccc gacggagcga tgtcgcctta ccaggccgcc 240 atgcacgatg gctccggata cgtgatgcat cgttcaatgc agtttgagga tggtgccatg 300 ctgcattcag accaccgcta catctataag ggaaaccata tcaaaggaga gtttcggctg 360 accggaagcg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgcgtcg acaagctgct gtacccaaac gacaacacca taatcggcaa attcgactgg 480 acctacacca ctaccagtgg caagcgctac caaagtgatg tgcagaccaa cgtcacattt 540 ggcaagccaa tagcggccga cattttgaag aagcagccaa tgttcgtgtt ccgcatgatg 600 gaactcaagc acaccaagac tgagctcaac ttcaagcagt ggcagaaggc attccaggac 660 atcgcctgat gcgctcgatc ctcaagtgta ttacattttg cttgacaaca cctcttagaa 720 atatcctttt tttrttctgtc aatacccaat attgcaacaa gaagatacaa gctttgtctc 780 agcagaactt tatctaaaca gtgtcgttct agaactgata atgatgggct cctacctttt 840 tatttttcaa ttaaaacttt ttaaaaatgt gctttatcta cgaaaatcca atgtatttac 900 atcattccgc caaacatttt ttctgggcaa ttaccgcttc ctcgatcata cttatgttct 960 cagatgggac acctcactca agagtataca tttagatcac tgcctcccca tattccgatt 1020 tatttttcat cttcaagggg gtgctggtaa tagtcacatg taaattctta cagtgaaatt 1080 ttttccatcg gtttctacca cttggcgttt tggaatgcat aggaactcgg aatgcctgca 1140 tttcttaacg tattgtaatt acaattacgg actacgtttt attcatcacg acgaaagaat 1200 ctctgcccat atggaaaaaa cccttccttc tcttgaaat 1239

<210> 56 <211 >874 <212> DNA <213> Branchiostoma lanceolatum <400> 56 acaaacatgc ctcttccagc gacccacgat ttacacatct ccggctctat caatggacat 60 gagtttgact tggagggcag tggcaagggc aatgcaaaag aaggttatca ggagctccac 120 ctaaagtcca acaggggtga cctgtcattc tccccctgga tcctggtccc aaacatcggc 180 tacggcttct accagtacct gcccttcccc gacggagcga tgtcgcctta ccaggccgcc 240 atgcacgatg getccggcta cgtgatgcat cgtacaatgc aatttgagga tggtgccatg 300 ctgcattcag accaccgcta tacctataag ggaaaccata tcaaaggaga gtttaggctg 360 accggaagtg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgcgtgg acaagctgct gtatcctact gagaacaccc taatcggcaa attcgactgg 480 acttacacca ctaccagcgg caagcgctac caaagtgatg tgcagaccaa cgtcaccttt 540 gccaagccaa tggctgccga cattctgaag aagcagccga tgttcgtgtt tcgcaaggtc 600 gaactcaagc acaccaagac agagctcaac ttcaagcagt ggcagaaggc attccaggac 660 atcgtgtgat gcgctcgatc ctccagtgta ttacatttgc tttacaacac cctaagaaat 720 atccatattt ttctgttaat accaaatatt tcaataagaa tatacaaact ttgtctcggc 780 agaacttaat caaaacagag tcgttctaga acttagaata atggtctcct accatttttt 840 tttcaatcga aactttttaa aaatgtgctt tgtt 874

<210> 57 <211 >961 <212> DNA <213> Branchiostoma lanceolatum <400> 57 acaaccatgt ctctcccagc gacccacgat ttacacatct ccggctcaat caatggacat 60 gagtttgact tggaaggcag tggcaagggc aatgcaaaag aaggttatca ggagctccac 120 ctaaagtcca acaagggtga cctgtcattc tccccctgga tcctggtccc aaacatcggc 180 tacggcttct accagtacct gcccttcccc gacggagcga tgtcgcctta ccaggccgcc 240 atgcacgatg gctccggata cgtgatgcat cgttcaatgc agtttgagga tggtgccatg 300 ctgcattcag accaccgcta catctataag ggaaaccata tcaaaggaga gtttcggctg 360 accggaagcg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgcgtcg acaagctgct gtacccaaac gacaacacca taatcggcaa attcgactgg 480 acctacacca ctaccagtgg caagcgctac caaagtgatg tgcagaccaa cgtcacattt 540 ggcaagccaa tagcggccga cattttgaag aagcagccaa tgttcgtgtt ccgcaaggtg 600 gaactcaagc acaccaagac tgagctcaac ttcaagcagt ggcagaaggc attccaggac 660 atcgcctgat gcgctcgatc ctcaagtgta ttacatttgc ttgacaacac ccttagaaat 720 atcctttttt ttttctgtca atacccaata ttgcaacaag aagatacaaa ctttgtctca 780 gcagaacttt atcaaaacag tgtcgttcta gaactgataa tgatggtctc ctaccatttt 840 attttttcaa ttaaaacttt ttaaaaatgt gctttatcta cgaaaatcca atgtattttc 900 accattcgtc aaacattttt ttcctgggcc attaccgctt ctcgatcata cttatgtctc 960 a 961

<210> 58 <211> 1712 <212> DNA <213> Branchiostoma lanceolatum <400> 58 acaaccatgt ctctcccagc gacccacgat ttacacatct ccggctctat caatggacat 60 gagtttgact tggagggcag tggcaagggc aatgcaaaag aaggttatca ggagctccac 120 ctaaagtcca acaagggtga cctgtcattc tccccctgga tcctggtccc aaacatcggc 180 tacggcttct accagtacct gcccttcccc gacggagcga tgtcgcctta ccaggccgcc 240 atgcacgatg gctccggcta cgtgatgcat cgtacaatgc aatttgagga tggtgccatg 300 ctgcattcag accaccgcta tacctataag ggaaaccata tcaaaggaga gtttaggctg 360 accgggagcg gtttccctgc tgacggccct gtgatgacca actcgctgac cgcggtggac 420 tggtgtgtgg ataagctgct gtaccccaac gagaacacca taatcggcaa attcgactgg 480 acctacacca ctaccagtgg caagcgctac caaagtgatg tgcagaccaa cgtcaccttt 540 gccaagccaa tagcggccga cattctgaag aagcagccga tgttcgtgtt ccgcaaggtg 600 gagctcaagc actccaagac cgagctcaac ttcaagcagt ggcagaaggc attccaggac 660 atcgtgtgat gcgctcgatc ctccagtgta ttacatttgc tttacaacat cctaagaaat 720 atccattttt ttctgttaat acccaatatt gcaacaagaa gatacaaact ttgtctcggc 780 agaacttaat caaaacagtg tcgttctaga acctagaata atggtctcct accatttttt 840 ttaaatcaaa actttttaaa aatgtgcttt gtttacgaat atctaatgta tttcaccatt 900 cgtcaaacat ttttttttcc tgggccatta ccgcttctcg ataatacttt tgttctcaag 960 atggacacct cactctagag tatacattca gatactgatt tcccatatcc gtttctttta 1020 atttcaaagg gtgttagtaa tcgtcaaatg taaatctaac tggatatttt ttatcgtttc 1080 taccactttg ctttttggta tgcatagaga gctgaatgca tgcatttctt atcgtatgta 1140 ttacaaatac ttctgacctt cccgttttga aattaacaga tggatatgta ctatcgacaa 1200 tcctctctct ctctatagga tcaaatcaca tgctattgcc taatcgggaa agatgcagta 1260 gctgcacctt cgtagcttca catctatttg atacaatcta aattccaaat atatgagctc 1320 tgtttttaaa ttcgctcttg ggataacagt aaagcctgtc ggaaatccgc gtgtggttgt 1380 agaaaaatga ggctagctgt cggcgtattt ggaaaagggt cttcaaattg aatgcgtttc 1440 ccattcagta ccatcttcag acagatcact tagtaattca ggtaataaag atctggttcg 1500 taaaaatgca agtataaaca tcggttgagc tcaaacaggg agattgtaga aatatatgcg 1560 aaatatcttg gaatatcact gaagcttttt agtttcaaaa gtagcctcga agacagccct 1620 atacccttag actgttcatt aaatttcttt tggtagttca gcttgaatac ataaagattg 1680 ggggtactaa aaacaaaaaa aaaaaaaaaa aa 1712

<210> 59 <211> 2155 <212> DNA <213> Branchiostoma lanceolatum <400> 59 acaatcatgt ctctcccagc aacccacgat ttacacatct ccggctcaat caatggacat 60 gagtttgact tggaaggcag tggcaagggc aatgcaaaag aaggttatca ggagctccac 120 ctaaagtcca acaagggtga cctgtcattc tccecctgga tcctggtccc aaacatcggc 180 tacggcttct accagtacct gcccttcccc gacggagcga tgtcgcctta ccaggccgcc 240 atgcacgatg gctccggata cgtgatgcat cgttcaatgc agtttgagga tggtgccatg 300 ctgcattcag aceaccgcta catctataag ggaaaccata tcaaaggaga gtttcggctg 360 accggaagcg gtttccctgc tgacggcect gtgatgacca actcgctgac c'gctgcggac 420 tggtgcgtcg acaagctgct gtacccaaac gacaacacca taatcggcaa attcgactgg 480 acctacacca ctaccagtgg caagcgctac caaagtgatg tgcagaccaa cgtcacattt 540 ggcaagccaa tagcggccga cattttgaag aagcagccaa tgttcgtgtt ccgcaaggtg 600 gaactcaagc acaccaagac tgagctcaac ttcaagcagt ggcagaaggc attccaggac 660 atcgcctgat gcgctcgatc ctcaagtgta ttacatttgc ttgacaacac ccttagaaat 720 atcctttttt tttctgtcaa tacccaatat tgcaacaaga agatacaaac tttgtctcag 780 cagaacttta tcaaaacagt gtcgttctag aactgataat gatggtctcc taccatttta 840 ttttttcaat taaaactttt taaaaatgtg ctttatctac gaaaatccaa tgtattttca 900 ccattcgtca aacatttttt cctggccatt accgcttctc gatcatactt atgttctcaa 960 gatggacacc tcactctaga gtatacattt agatactgct tcccatatcc gtttcttttc 1020 atttcaaggg gtgttggtaa tagtcaaatg taaatcttgc aggatatttt tcategtttt 1080 taccactttg ctttttggta tgccaggaca tcgcctgatg cgctcgatcc tcaagtgtat 1140 tacatttgct ggacaacacc cttagaaata tcettttttt ttcgttcaat accccatatt 1200 gcaacaagaa gatacaaact ttgtctcagc agaactttat caaaacagtg tcgttctaga 1260 actgataatg atggtctcct accattttat tttttcaatt aaaacttttt aaaaatgtgc 1320 tttatctacg aaaatccaat gtattttcac cattcgtcaa acattttttc ctggccatta 1380 ccgcttctcg atcatactta tgttctcaag atggacacct cactctagag tatacattta 1440 gatactgctt cccatatccg tttcttttca tttcaagggg tgttggtaat agtcaaatgt 1500 aaatcttgca ggatattttt catcgttttt accactttgc tttttggtat gcatagagaa 1560 ctgaatgcat gcatttctta tcgtatgtaa tacaatacag accttcctgt tttattatca 1620 gcagatagat ctgcactatg aacaatcctc tctctagata taggatcaaa ttacatgcta 1680 ttgcctaatc aggtagatac agtagctgac cccccccgta gcttcaaatc tatatgatac 1740 aaatcaaaat tcaaaatata tgagctctgt ttttaaattc gctcttggta taacaataaa 1800 gtctgtcgga aacccgcgtg tggttgtaag aaaacgaggt taactgtcgg cgtattttac 1860 aaatttaatg cgtttcccat taagtccctt tttcagaaag atcacttagt aattcaggta 1920 gtaaagatct ggttcgtaaa aatgttatga taaatatcgg ttaagctcaa aaagggtgat 1980 tgtagatata tatgcgaaat atcttcgaat atcactgaag ccttttagtt tcaaaagtcg 2040 cctccaagac aggcctatat cctacggctg ttcattaaac ttctttttgt ggttcaactt 2100 gaagaaataa aaagatgagg gtacttttaa aaaaaaaaaa aaaaaaaaaa aaaaa 2155

<210> 60 <211 >721 <212> DNA <213> Branchiostoma lanceolatum <400> 60 aacaacatgt ctctccctaa gacccacgat ttacacatct ccggctctgt caatggacat 60 gagtttgact tggagggcag tggcaagggc aatgcaaaag aaggttatca ggagctccac 120 ctaaagtcca acaggggtga cctgtcattc tccccttgga tcctggtccc aaacatcggc 180 tatggcttct accagtacct gcccttcccc gacggagcga tgtcgcctta ccaggccgcc 240 atgcacgatg gctccggata cgtgatgcat cgtgcaatgc ggtttgagga tggtgctatg 300 ctgcattcag accaccgcta tacctacaac ggaaacaata tcaaaggaga gtttcggctg 360 accgggagcg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgtgtgg acaagctget gtaccccaac gagaacacca ttatcggcaa attcgactgg 480 acatacacca ctacaagtgg caagcgctac caaagtgatg tgcagaccaa cgtcaccttt 540 ggcaagccaa tatcggccga cattctgaag aagcagccga tgttcgtgtt ccgtaaggtg 600 gaactcaagc actccaagac cgagctcaac ttcaagcagt ggcagaaggc attccaggac 660 atcgtgtgat gcgctcgatc ctccagtgta ttacatttgc tttacaacac cctaaaaaaa 720 a 721

<210> 61 <211> 925 <212> DNA <213> Branchiostoma lanceolatum <400> 61 acaaacatgc ctcttccagc gacccacgat ttacacatct ccggctctat caatggacat 60 gagtttgact tggagggcag tggcaagggc aatgcaaaag aaggttatca ggagctccac 120 ctaaaatcca acaagggtga cctgtcattc tccccctgga tcctggtccc aaacatcggc 180 tacggcttct accagtacct gcccttcccc gacggagcga tgtcgcctta ccaggccgcc 240 atgcacgatg gctccggata cgtgatgcat cgtgcaatgc ggtttgagga tggtgccatg 300 ctgcattcag accaccgcta tacctacaac ggaaaccata tcaaaggaga gtttcggctg 360 accgggagcg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgtgtgg ataagctgct gtaccccgac gagaacacca ttatcggcaa attcgactgg 480 acatacacca ctaccagtgg caagcgctac caaagtgatg tgcagaccaa cgtcacattt 540 gccaagccaa tatcggccga cattctgaag aagcagccga tgttcgtgtt ccgtaaggtg 600 gagctcaagc actccaagac cgagctcaac ttcaagcagt ggcaaaaggc attccaggac 660 atcgtgtgat gcgctcgatc ctccagtgta ttacatttgc tttacaacac cctaagaaat 720 ttttttttct gttaataccc aatattgcaa caagaagata caaacattgt attggcagaa 780 gttaatcaaa acagtgtcgt tctagaacta agaatgatgg tctcctatca ttttattttt 840 caatcaaaac tttttaaaat tgtgctttgt ttacgaatat ccaatctata ttttcaccat 900 tcgtcaaaca ttttttcctg gccat 925 <210> 62 <211 >926

<212> DNA <213> Branchiostoma lanceolatum <400> 62 acaaccatgt ctctcccagc gacccacgat ttacacatct ccggctctat caatggacat 60 gagtttgact tggagggcag tggcaagggc aatgcaaaag aaggttatca ggagctccac 120 ctaaaatcca acaagggtga cctgtcattc tccccctgga tcctggtccc aaacatcggc 180 tacggettct accagtacct gcccttcccc gacggagcga tgtcgcctta ccaggccgcc 240 atgcacgatg gctccggata cgtgatgcat cgtgcaatgc ggtttgagga tggtgccatg 300 ctgcattcag accaccgcta tacctacaac ggaaaccata tcaaaggaga gtttcggctg 360 accgggagcg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgtgtgg araagctgct gtaccccgac gagaacacca ttatcggcaa attcgactgg 480 acatacacca ctaccagtgg caagcgctac caaagtgatg tgcagaccaa cgtcacattt 540 gccaagccaa tatcggccga cattctgaag aagcagccga tgttcgtgtt ccgtaaggtg 600 gagctcaagc actccaagac cgagctcaac ttcaagcagt ggcaaaaggc attccaggac 660 atcgtgtgat gcgctcgatc ctccagtgta ttacatttgc tttacaacac cctaagaaat 720 ttttttttct gttaataccc aatattgcaa caagaagata caaacattgt attggcagaa 780 gttaatcaaa acagtgtcgt tctagaacta agaatgatgg tctcctatca ttttattttt 840 caatcaaaac tttttaaaat tgtgctttgt ttacgaatat ccaatctata ttttcaccat 900 tcgtcaaaca tttttttcct ggccat 926

<210> 63 <211 >726 <212> DNA <213> Branchiostoma lanceolatum <400> 63 aacaacatgt ctctccctaa gacccacgat ttacacatct ccggctctgt caatggacat 60 gagtttgact tggagggcag tggcaagggc gatgcaaaag aaggttatca ggagctccac 120 ctaaagtcca acaggggtga cctgtcattc tccccttgga tcctggtccc aaacatcggc 180 tatggcttct accagtacct gcccttcccc gacggagcga tgtcgcctta ccaggccgcc 240 atgcacgatg gctccggata cgtgatgcat cgtgcaatgc ggtttgagga tggtgctatg 300 ctgcattcag accaccgcta tacctacaac ggaaacaata tcaaaggaga gtttcggctg 360 accgggagcg gtttccctgc tgacggccct gtgatgacca actcgctgac cgctgcggac 420 tggtgtgtgg acaagctgct gtaccccaac gagaacacca ttatcggcaa attcgactgg 480 acatacacca ctacaagtgg caagcgctac caaagtgatg tgcagaccaa cgtcaccttt 540 ggcaagccaa tatcggccga cattctgaag aagcagccga tgttcgtgtt ccgtaaggtg 600 gaactcaagc actccaagac cgagctcaac ttcaagcagt ggcagaaggc attccaggac 660 atcgtgtgat acgctcgatc ctccagtgta ttacatttgc tttacaacac cctaaaaaat 720 aaaaaa 726

<210> 64 <211 >598 <212> DNA <213> Branchiostoma lanceolatum <400> 64 acaaccatgt ctctcccagc gacccatgag ttgcacattt ttgacaaaat caatggccat 60 gagtttgaca tgaggggtaa aggcaccggt aacccaaatg .acggttatga ggaccttgac 120 ctgaagtcca ccaaggatga ccttccattc tccccctgga tcctggtcca aaacatcggg 180 tacggcttta accagtacct gccctacccc gacggagcga tgtcgccttt tcaggctgcc 240 atgtacaatg gctccgggta ccacgtccat cgtgaaatgg ggtttgaaga cggtgccacg 300 gtgactggca tctaccgcta cacctacgag ggaagccaca tcaaaggaga gtttcaggtg 360 gatgggaccg gattccctgc tgacggccct gtgatgacca actcgctcac tgatcaggac 420 tggtccgtga ccaagatgat gtaccttgat aacaaaaccg tcactagcac cgctgaccag 480 acctacacca ctgcaagtgg caagcgctac cagggcacag tgcggaccaa caacaccttt 540 gccaagccga tagcggccaa catcctgcag aagcagccgg tattcgtgtc ccgcaagc 598

<210> 65 <211 >595 <212> DNA <213> Branchiostoma lanceolatum <400> 65 aacaacatgt ctctccctaa gacccacgag ttgcatattt ttggcaaaat caatggccat 60 gagtacgaca tgaggggtaa aggcacaggt aacccaaatg acggttatga ggaccttgac 120 ctgaagtcca aggatgacct tccattctcc ccctggatcc tggttcaaaa catcggatac 180 ggctttaacc agtacctgcc ctaccccgac ggagcgatgt cgcctttcca ggctgccatg 240 tgcgatggct ccgggtacga ggtccatcgt gaaatggagt ttgaagacgg tgccacgctg 300 actggcatct accgctacac ctacgaggga agccacatca aaggagagtt tcaggtggat 360 gggaccggtt tccctgatga cggccctgtg atgaccgact cgctcaccga tctggactgg 420 gtcgtgacca agatggtgta tcccgacgag aaaaccgtct tcagcacctc cgaccagacc 480 tacaccactg caagtggcaa gggttacaag agtacagtgc ggaccaacaa catttttgcc 540 aagccaatgg cggccgacat gatgcagaac cagccgatat tcgtgtcccg caagc 595

<210> 66 <211 >598 <212> DNA <213> Branchiostoma lanceolatum <400> 66 acaactatgt ctctcccagc gacccatgag ttgcacattt ttggcaaaat caatggccat 60 gagtacgaca tgaggggtaa aggcactggt aacccaaatg acggttatga ggaccttgac 120 ctgaagtcca ccaaggatga ccttccattc tccccctgga tcctggtcca aaacatcggg 180 tacggettta accagtactt gccctacccc gacggagcga tgtcgccttt ccaggctgcc 240 atgtgcgatg gctccgggta cgaggtccat cgtgaaatgg agtttgaaga cggtgccacg 300 gtgactggta tctaccgcta cacctacgag ggaagccaca tcaaaggaga gtttcaggtg 360 gatgggaccg gtttccctga tgacggccct gtgatgaccg actcgctcac tgatctggac 420 tgggtcgtga ccaagatggt gtaccccgac gagaaaaccg tcttcagcac ctccgaccag 480 acctacacca ctacaagtgg caagggctac aagagcacag tgcggaccaa caacattttt 540 gccaagccaa tagcggccga catgatgcag agccagccgg tattcgtgtc ccgcaagc 598

<210> 67 <211 >598 <212> DNA <213> Branchiostoma lanceolatum <400> 67 acaaccatgt ctctcccagc gacccatgag ttgcacattt ttggcaaaat caatggccat 60 gagtttgaca tgaggggtaa aggcaccggt aacccaaatg acggttatga ggaccttgac ' 120 ctgaagtcca ccaaggatga ccttccattc tccccctgga tcctggtcca aaacatcggg 180 tacggettta accagtacct gccctacccc gacggagcga tgtcgccttt tcaggctgcc 240 atgtacaatg gctccgggta ccacgtccat cgtgaaatgg agtttgaaga cggtgccacg 300 ctgactggca tctaccgcta cacctacgag ggaagccaca tcaaaggaga gtttcaggtg 360 gatgggaccg gtttccctgc tgacggccct gtgatgaccg actcgctcac tgatctggac 420 tgggtcgtga caaagatggt gtatcccgac gacaaaaccg tcttcagcac ctctgaccag 480 acctacacca ctacaagtgg caagggctac cagagcacag tgcggaccaa caacattttt 540 gccgagccga tagcggccga catgatgcag agccagccgg tattcgtgtc ccgcaagc 598

<210> 68 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 68

Met Pro Leu Pro Ala Thr His Glu Leu His Ile Phe Gly ser lie Asn 1 5 10 15

Ser Leu Glu Phe Asp Leu val Gly Arg Gly Thr Gly Asn Pro Lys Glu 20 25 30

Gly Tyr Glu Glu Leu His Leu Lys Ser Thr Lys Ser Ala Leu Gin Phe 35 40 45 ser Pro Trp lie Leu Val Pro Gin.ile Gly Tyr Gly Phe Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met Ser Pro Phe Gin Ala Ala Met Asn 65 70 75 80

Asp Gly ser Gly Tyr Gin val His Arg Thr Met Gin Phe Glu Asp Gly 85 90 95

Ala Thr Leu Thr Gly lie Tyr Arg Tyr Thr Tyr Glu Gly Thr His lie 100 105 110

Lys Gly Glu Phe Gin val lie Gly Thr Gly Phe Pro Ala Asp Gly Pro H5 120 125

Val Met Thr Asn ser Leu Thr Ala Ala Asp Trp Cys val Thr Lys lie 130 135 140 val Tyr Pro Asn Glu Asn Thr He lie Asp Lys Phe Asp Trp Thr Tyr

145 150 H 155 lBO

Thr Thr Thr Ser Gly Lys Arg Tyr Gin Ser Asn Val Arg Ser Asn Phe 165 170 175

Thr Phe Ala Lys Pro He Ala Ala Asn He Leu Gin Lys Gin Pro Met 180 185 190

Phe val Phe Arg Lys Thr Glu Leu Lys His ser Lys Thr Glu Leu Asn 195 200 205

Phe Lys Glu Trp Gin Thr Ala phe Ser Asp val Met 210 215 “ 220

<210> 69 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 69

Met ser Leu Pro Ala' Thr His Glu Leu His Ile Phe Giv ser Tie Asn 1 5 10 15 ser Leu Glu Phe Asp Leu Val Gly Arg Gly Thr Gly Asn Pro Arq Glu 20 25 30

Gly Tyr Glu Glu Leu His Leu L^s ser Thr Lys ser Ala Leu Gin Phe

Ser Pro Trp lie Leu val Pro Gin lie Gly Tyr Gly phe Tyr Gin Tvr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met ser pro Phe Gin Ala Ala Met Asn 65 70 75 80

Asp Gly ser Gly Tyr Gin Val His Arg Thr Met Gin Phe Glu His gIv 85 90 95

Ala Thr Leu Thr Gly lie Tyr Arg Tyr Thr Tyr Glu Gly Thr His lie 100 105 110

Lys Gly Glu Phe Gin val lie Gly Thr Gly Phe Pro Ala Asp Gly pro 115 120 125

Val Met Thr Asn ser Leu Thr Ala Ala Asp Trp cys val Thr Lys lie 130 135 140

Val Tyr Pro Asn Glu Asn Thr lie lie Asp Lys Phe Asp Trp Thr Tyr 145 150 155 1§0

Thr Thr Thr ser Gly Lys Arg Tyr His ser Asn val Arg ser Asn phe 165 170 175

Thr Phe Ala Lys Pro lie Ala Ala Asn lie Leu Gin Lys Gin Pro Met 180 185 190

Phe val Phe Arg Lys Thr Glu Leu Lys His Ser Lys Thr Glu Leu Asn 195 200 205

Phe Lys Glu Trp Gin Thr Ala Phe Gly Asp val Met 210 215 220

<210> 70 <211 > 219 <212> PRT <213> Branchiostoma lanceolatum <400> 70

Met ser Leu Pro Ala Thr His Glu Leu His lie Phe Gly ser Phe Asn 15 10 15

Gly val Asp Phe Asp Met Val Gly Arg Gly Thr Gly Asn Pro Asn Asp 20 25 30

Gly Tyr Glu Glu Leu Asn Leu Lys ser Thr Lys Gly Ala Leu Gin Phe 35 40 45 ser Pro Trp Ile Leu Val Pro Gin Ile Gly Tyr Gly Phe His Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Met ser Pro Phe Gin Ala Ala Met Lys Asp 65 70 75 80

Gly ser Gly Tyr Gin val His Arg Thr Met Gin Phe Glu Asp Gly Ala 85 90 95 ser Leu Thr Ser Asn Tyr Arg Tyr Thr Tyr Glu Gly ser His ile Lys 100 105 110

Gly Glu phe Gin val ile Gly Thr Gly Phe pro Ala Asp Gly Pro Val 115 120 125.

Met Thr Asn Ser Leu Thr Ala Ala Asp Trp cys Val Thr Lys Met Leu 130 135 140

Tyr pro Asn Asp Lys Thr Ile ile Ser Thr Phe Asp Trp Thr Tyr Thr 145 150 155 160

Thr Gly ser Gly Lys Arg Tyr Gin Ser Thr val Arg Thr Asn Tyr Thr 165 170 ' 175

Phe Ala Lys pro Met Ala Ala Asn ile Leu Lys Asn Gin Pro Met Phe 180 185 190 val phe Arg Lys Thr Glu Leu Lys His ser Lys Thr Glu Leu Asn Phe 195 200 205

Lys .Glu Trp Gin Lys Ala Phe Thr Asp val Met 210 215

<210> 71 <211> 219 <212> PRT <213> Branchiostoma lanceolatum <400> 71

Met Pro Leu Pro Ala Thr His Glu Leu His ile Phe Gly ser Phe Asn 15 10 15

Gly val Asp phe Asp Met val Gly Arg Gly Thr Gly Asn Pro Asn Asp 20 25 30

Gly Tyr Glu Glu Leu Asn Leu Lys ser Thr Lys Gly Ala Leu Gin Phe 35 40 45

Ser Pro Trp Ile Leu Val Pro Gin ile Gly Tyr Gly Phe His Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Met ser pro Phe Gin Ala Ala Met Lys Asp 65 70 75 80

Gly ser Gly Tyr Gin Val His Arg Thr Met Gin Phe Glu Asp Gly Ala 85 90 95 ser Leu Thr ser Asn Tyr Arg Tyr Thr Tyr Glu Gly ser His ile Lys 100 105 110

Gly Glu Phe Gin Val ile Gly Thr Gly Phe Pro Ala Asp Gly Pro Val 115 120 125

Met Tao Asn Ser LeU Τ^Γ ΤτΓ A^a Asp Trp cys val Thr Lys Met Leu 130 135 140

Tyr Pro Asn Asp Lys Thr ile ile ser Thr Phe Asp Trp Thr Tyr Asn 145 150 155 160

Thr Ala Ser Gly Lys Arg Tyr Gin ser Thr val Arg Thr Asn Tyr Thr 165 170 175

Phe Asn Lys Pro Met Ala Ala Asn ile i_eu Lys Asn Gin Pro Met Phe 180 185 190

Val Phe Arg Lys Thr Glu Leu Lys His ser Lys Thr Glu Leu Asn Phe 195 200 205

LyS 210 ΤΓΡ Gln M6t AU 215 AU ASP Va1 Met

<210> 72 <211 > 219 <212> PRT <213> Branchiostoma lanceolatum <400> 72

Met pro Leu Pro Ala Thr His Glu Leu His ile Phe Gly ser Phe Asn 1 5 10 15

Gly Val Asp Phe Asp Met Val Gly His Gly Thr Gly Asn Pro Asn Asp 20 25 30

Gly Tyr Glu Glu Leu Asn Leu Lys Ser Thr Lys Gly Ala Leu Gin Phe 35 40 45 ser Pro Trp ile Leu Val Pro Gin ile Gly Tyr Gly Phe His Gin Tyr 50 55 y 60

Leu Pro phe Pro Asp Gly Met ser Pro Phe Gin Ala Ala Met Lys Asp 65 70 75 80

Gly Ser Gly Tyr Gin val His Arg Thr Met Gin Phe Glu Asp Gly Ala 85 90 95 ser Leu Thr ser Asn Tyr Arg Tyr Thr Tyr Glu Gly ser His ile Lys 100 105 110

Gly Glu Phe Gin Val Asn Gly Thr Gly Phe Pro Ala Asp Gly pro val 115 120 125

Met Thr Asn Ser Leu Thr Thr Ala Asp Trp cys val Thr Lys Met Leu 130 135 140

Tyr Pro Asn Asp Lys Thr ile Ile ser Thr Phe Asp Trp Thr Tyr Thr 145 150 155 160

Thr Gly ser Gly Lys Arg Tyr Gin Ser Thr Val Arg Thr Asn Tyr Thr 165 170 175

Phe Ala Lys pro Met Ala Ala Asn ile Leu Gin Asn Gin Pro Met Phe 180 185 190 val Phe Arg tys Thr Glu Leu Lys His ser Lys Thr Glu Leu Asn Phe 195 200 205

Lys Glu Trp Gin Lys Ala Phe Ala Asp val Met 210 215

<210> 73 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 73

Met ser Leu Pro Ala Thr His Glu Leu His ile Phe Gly ser Ile Asn 1 5 10 15

Ser Leu Glu Phe Asp Leu val Gly Arg Gly Thr Gly Asn Pro Lys Glu 20 25 30

Gly Tyr Glu Glu Leu His Leu Lys Ser Thr Lys ser Ala Leu Gin Phe 35 40 45

Ser Pro Trp ile Leu val Pro Gin Ile Gly Tyr Gly Phe Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met Ser pro Phe Gin Ala Ala Met Asn 65 70 75 80

Asp Gly ser Gly Tyr Gin Val His Arg Thr Met Gin Phe Glu Asp Gly 85 90 95

Ala Thr Leu Thr Gly ile Tyr Arg Tyr Thr Tyr Glu Gly Thr His ile 100 105 110

Lys Gly g|u Phe Gin Val ile Gig Thr Gly Phe pro Ala Asp Gly Pro val Met Thr Asn Ser Leu Thr Ala Ala Asp Trp cys val Thr Lys ile 130 135 140

Val Tyr Pro Asn Glu Asn Thr Ile ile Asp Lys Phe Asp Trp Thr Tyr ±4:> 150 155 150

Thr Thr Thr ser Gly Lys Arg Tyr Gin ser Asn Val Arg ser Asn Phe 165 170 175

Thr phe Ala Lys Pro ile Ala Ala Asn ile Leu Gin Lys Gin Pro Met 180 185 190

Phe val Phe Arg Lys Thr Glu Leu Lys His ser Lys Thr Glu Leu Asn 195 200 205

Phe Lys Glu Trp Gin Thr Ala Phe Ser Asp Val Met 210 215 220

<210> 74 <211 > 219 <212> PRT <213> Branchiostoma lanceolatum <400> 74

Met Pro Leu Pro Ala Thr His Glu Leu His Ile Phe Gly Ser Phe Asn 1 5 10 15

Gly Val Glu Phe Asp Met Val Gly Arg Gly Thr Gly Asn pro Asn Asp 20 25 30

Gly Ser Glu Asp Leu His Leu Lys ser Thr Lys Gly Ala Leu Gin Phe 35 40 45 ser Pro Trp ile Leu ile Pro His Ile Gly Tyr Gly phe His Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Met ser Pro Phe Gin Ala Ala Met Gin Asp 65 70 75 80

Gly ser Gly Tyr Gin val His Arg Thr Met Gin phe Glu Asp Gly Ala 85 90 95 ser Leu Thr Ala Asn Phe Arg Tyr Thr Tyr Glu Gly ser His Ile Lys 100 105 110

Gly Glu Phe Gin Val ile Gly ser Gly phe Pro Ala Asp Gly Pro Val 115 120 125

Met Thr Asn ser Leu Thr Ala Val Asp Trp Cys Val Ala Lys Met Leu 130 135 140

Tyr Pro Asn Asp Lys Thr ile ile Ser Thr Phe Asp Trp Thr Tyr Thr 145 150 155 160

Thr Gly ser Gly Lys Arg Tyr Gin Ser Thr Val Arg Thr Asn Tyr Thr 165 170 175 phe Ala Lys Pro Met Ala Ala Asn Ile Leu Lys Asn Gin Pro Met Phe 180 185 190 val Phe Arg Lys Thr Glu Leu Lys His ser Lys Thr Glu Leu Asn Phe 195 200 205

Lys Glu Trp Gin Lys Ala Phe Ala Asp val Met 210 215

<210> 75 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 75

Met Pro Leu Pro Ala Thr His Asp Leu His Ile ser Gly Ser lie Asn 1 5 10 15

Gly His Glu Phe Asp Leu Glu Gly ser Gly Lys Gly Asn Glu Lys Glu 20 25 30

Gly Tyr Gin Glu Leu His Leu Lys ser Asn Lys Gly Asp Leu ser Phe 35 40 45 ser Pro Trp lie Leu Val Pro Asn Thr Gly Tyr Gly Phe Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met ser Pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly Ser Gly Tyr Val Met His Arg Ser Met Gin Phe Glu Asp Gly 85 90 95

Ala Met Leu His ser Asp His Arg Tyr lie Tyr Lys Gly Asn His lie 100 105 110

Lys Gly Glu Phe Arg Leu Thr Gly ser Gly Phe Pro Ala Asp Gly Pro 115 120 125 val Met Thr Asn Ser Leu Thr Ala Ala Asp Trp cys val Asp Lys Leu 130 135 140

Leu Tyr Pro Asn Asp Asn Thr lie lie Gly Lys Phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr ser Gly Lys Arg Tyr Gin Ser Asp val Gin Thr Asn Val 165 170 175

Thr Phe Gly Lys Pro lie Ala Ala Asp lie Leu Lys Lvs Gin Pro Met 180 185 190

Phe Val Phe Arg Met Met Glu Leu Lys His Thr Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp lie Ala 210 215 220

<210> 76 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 76

Met Pro Leu Pro Ala Thr His Asp Leu His lie Ser Gly Ser lie Asn 1 5 10 15

Gly His Glu Phe Asp Leu Glu Gly Ser Gly Lys Gly Asn Ala Lys Glu 20 25 30

Gly Tyr Gin Glu Leu His Leu Lys ser Asn Arg Gly Asp Leu ser Phe 35 40 45 ser Pro Trp ile Leu val Pro Asn ile Gly Tyr Gly Phe Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met ser Pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly Ser Gly Tyr Val Met His Arg Thr Met Gin Phe Glu Asp Gly 85 90 95

Ala Met Leu His Ser Asp His Arg Tyr Thr Tyr Lys Gly Asn His Ile 100 105 110

Lys Gly Glu Phe Arg Leu Thr Gly ser Gly Phe pro Ala Asp Gly pro 115 120 125 val Met Thr Asn ser Leu Thr Ala Ala Asp Trp cys val Asp Lys Leu 130 135 140

Leu Tyr Pro Thr Glu Asn Thr Leu Ile Gly Lys Phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr s er Gly Lys Arg Tyr Gin Ser Asp val Gin Thr Asn Val 165 170 175

Thr Phe Ala Lys Pro Met Ala Ala Asp Ile Leu Lys Lys Gin Pro Met 180 185 190

Phe Val Phe Arg Lys val Glu Leu Lys His Thr Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp ile Val 210 215 220

<210> 77 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 77

Met ser Leu Pro Ala Thr His Asp Leu His ile ser Gly ser ile Asn 15 10 25

Gly His Glu Phe Asp Leu Glu Gly ser Gly Lys Gly Asn Ala Lys Glu 20 25 30

Gly Tyr Gin Glu Leu His Leu Lys ser Asn Lys Gly Asp Leu ser Phe 3 5 40 4 5 ser pro Trp ile Leu Val Pro Asn Ile Gly Tyr Gly Phe Tyr Gin Tyr 50 55 50

Leu pro Phe Pro Asp Gly Ala Met ser Pro Tyr dn Ala Ala Met His bi> 70 75 80 asd Giv Ser Gly Tyr Val Met His Arg Ser Met Gin Phe Glu Asp Gly A5p y 7 85 90 95

Ala Met Leu His ser Asp His Arg Tyr ile Tyr Lys Gly Asn His ile 100 105 110

Lvs Gly Glu Phe Arg Leu Thr Gly Ser Gly Phe Pro Ala Asp Gly Pro 115 120 125 val Met Thr Asn ser Leu Thr Ala Ala Asp Trp cys val Asp Lys Leu 130 - 135 140

Leu Tyr Pro Asn Asp Asn Thr ile ile Gly Lys Phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr ser Gly Lys Arg Tyr Gin ser Asp val Gin Thr Asn Val 165 170 175

Thr Phe Gly Lys Pro ile Ala Ala Asp ile Leu Lys Lys Gin Pro Met 180 185 190

Phe val Phe Arg Lys Val Glu Leu Lys His Thr Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp ile Ala 210 215 220

<210> 78 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 78

Met ser Leu Pro Ala Thr His Asp Leu His ile ser Gly Ser ile Asn 1 5 10 15

Gly His Glu Phe Asp Leu Glu Gly ser Gly Lys Gly Asn Ala Lys Glu 20 25 30

Gly Tyr Gin Glu Leu His Leu Lys ser Asn Lys Gly Asp Leu ser Phe 35 40 J 45 ser Pro Trp ile Leu val Pro Asn Ile Gly Tyr Gly Phe Tyr Gin Tvr , 50 55 60

Leu Pro phe pro Asp Gly Ala Met ser Pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly ser Gly Tyr Val Met His Arg Thr Met Gin Phe Glu Asp Gly 85 90 95K 7

Ala Met Leu His Ser Asp His Arg Tyr Thr Tyr Lys Gly Asn His Ile 100 105 110

Lys Gly Glu Phe Arg Leu Thr Gly ser Gly Phe Pro Ala Asp Giv Pro 115 120 125 val Met Thr Asn ser Leu Thr Ala Val Asp Trp cys val Asp Lys Leu 130 135 140

Leu Tyr Pro Asn Glu Asn Thr ile ile Gly Lys phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr ser Gly Lys Arg Tyr Gin ser Asp val Gin Thr Asn val 165 170 175

Thr Phe Ala Lys Pro Ile Ala. Ala Asp ile Leu Lys Lys Gin Pro Met 180 185 190

Phe val Phe Arg Lys Val Glu Leu Lys His ser Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala phe Gin Asp ile val 210 215 220

<210> 79 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 79

Met ser Leu Pro Ala Thr His Asp Leu His Ile ser Gly ser ile Asn 15 10 15

Gly His Glu Phe Asp Leu Glu Gly Ser Gly Lys Gly Asn Ala Lys Glu 20 25 30

Gly Tyr Gin Glu Leu His Leu Lys ser Asn Lys Gly Asp Leu ser Phe 35 40 45 ser pro Trp ile Leu Val Pro Asn Ile Gly Tyr Gly Phe Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met Ser Pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly ser Gly Tyr Val Met His Arg ser Met Gin Phe Glu Asp Gly 85 90 95

Ala Met Leu His ser Asp His Arg Tyr ile Tyr Lys Gly Asn His ile 100 105 110

Lys Gly Glu Phe Arg Leu Thr Gly Ser Gly Phe Pro Ala Asp Gly Pro 115 120 125 val Met Thr Asn Ser Leu Thr Ala Ala Asp Trp Cys Val Asp Lys Leu 130 135 140

Leu Tyr Pro Asn Asp Asn Thr ile Ile Gly Lys Phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr ser Gly Lys Arg Tyr Gin ser Asp Val Gin Thr Asn val 165 170 175

Thr Phe Gly Lys Pro ile Ala Ala Asp ile Leu Lys Lys Gin Pro Met 180 185 190

Phe Val Phe Arg Lys val Glu Leu Lys His Thr Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala phe Gin Asp Ile Ala 210 215 220

<210> 80 <211> 220 <212> PRT <213> Branchiostoma lanceolatum <400> 80

Met ser Leu Pro Lys Thr His Asp Leu His Ile ser Gly ser val Asn 15 10 15

Gly His Glu Phe Asp Leu Glu Gly ser Gly Lys Gly Asn Ala lvs Glu 20 25 30

Gly Tyr Gin Glu Leu His Leu Lys ser Asn Arg Gly Asp Leu ser Phe 35 40 45 ser Pro Trp Ile Leu Val Pro Asn Ile Gly Tyr Gly Phe Tyr Gin Tvr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met ser Pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly ser Gly Tyr Val Met His Arg Ala Met Arg Phe Glu Asp Gly 85 90 95

Ala Met Leu His Ser Asp His Arg Tyr Thr Tyr Asn Gly Asn Asn ile 100 105 110

Lys Gly Glu Phe Arg Leu Thr Gly ser Gly Phe Pro Ala Asp Gly pro 115 120 125

Val Met Thr Asn ser Leu Thr Ala Ala Asp Trp cys val Asp Lys Leu 130 135 140

Leu Tyr Pro Asn Glu Asn Thr ile ile Gly Lys phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr ser Gly Lys Arg Tyr Gin ser Asp Val Gin Thr Asn val 165 170 175

Thr Phe Gly Lys pro ile Ser Ala Asp ile Leu Lys Lys Gin Pro Met 180 185 190

Phe Val Phe Arg Lys Val Glu Leu Lys His ser Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp ile val 210 215 220

<210> 81 <211> 220 <212> PRT <213> Branchiostoma lanceolatum <400> 81

Met Pro Leu pro Ala Thr His Asp Leu His ile ser Gly ser He Asn 1 5 10 15

Gly His Glu Phe Asp Leu Glu Gly ser Gly Lys Gly Asn Ala Lys Glu 20 25 30

Gly Tyr Gin Glu Leu His Leu Lgs ser Asn Lys Gly Asp Leu Ser Phe

Ser Pro Trp lie Leu Val Pro Asn He Gly Tyr Gly Phe Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met ser Pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly ser Gly Tyr Val Met His Arg Ala Met Arg Phe Glu Asp Gly 85 90 95

Ala Met Leu His ser Asp His Arg Tyr Thr Tyr Asn Gly Asn His lie 100 105 110

Lys Gly Glu phe Arg Leu Thr Gly ser Gly phe Pro Ala Asp Gly Pro 115 120 * 125 val Met Thr Asn ser Leu Thr Ala Ala Asp Trp cys Val Asp Lys Leu 130 135 140

Leu Tyr Pro Asp Glu Asn Thr lie lie Gly Lys Phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr Ser Gly Lys Arg Tyr Gin Ser Asp Val Gin Thr Asn Val 165 170 175

Thr Phe Ala Lys Pro lie ser Ala Asp lie Leu Lys Lys Gin Pro Met 180 185 J 190

Phe Val Phe Arg Lys Val Glu Leu Lys His ser Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp lie val 210 215 220

<210> 82 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 82

Met Ser Leu Pro Ala Thr His Asp Leu His lie ser Gly ser lie Asn 15 10 15

Gly His Glu Phe Asp Leu Glu Gly ser Gly Lys Gly Asn Ala Lys Glu 20 25 BO

Gly Tyr Gin Glu Leu His Leu Lys Ser Asn Lys Gly Asp Leu ser phe 35 40 45 ser Pro Trp Ile Leu Val Pro Asn Ile Gly Tyr Gly Phe Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met Ser pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly ser Gly Tyr Val Met His Arg Ala Met Arg Phe Glu Asp Gly 85 90 95

Ala Met Leu His ser Asp His Arg Tyr Thr Tyr Asn Gly Asn His ile 100 105 110

Lys Gly Glu Phe Arg Leu Thr Gly ser Gly phe Pro Ala Asp Gly Pro 115 120 125

Val Met Thr Asn ser Leu Thr Ala Ala Asp Trp cys val Asp Lys Leu 130 135 140

Leu Tyr Pro Asp Glu Asn Thr ile Ile Gly Lys Phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr Ser Gly Lys Arg Tyr Gin ser Asp Val Gin Thr Asn val 165 170 175

Thr Phe Ala Lys Pro Ile ser Ala Asp ile Leu Lys Lys Gin Pro Met 180 185 190 phe Val Phe Arg Lys Val Glu Leu Lys His ser Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp Ile Val 210 215 220

<210> 83 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <400> 83

Met Ser Leu Pro Lys Thr His Asp Leu His Ile Ser Gly Ser Val Asn 1 5 10 15

Gly His Glu Phe Asp Leu Glu Gly Ser Gly Lys Gly Asp Ala Lys Glu 20 25 30

Gly Tyr Gin Glu Leu His Leu Lys ser Asn Arg Gly Asp Leu ser Phe 35 40 45 ser pro Trp ile Leu val Pro Asn ile Gly Tyr Gly Phe Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met Ser Pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly ser Gly Tyr Val Met His Arg Ala Met Arg Phe Glu Asp Gly 85 90 95

Ala Met Leu His ser Asp His Arg Tyr Thr Tyr Asn Gly Asn Asn lie 100 105 110

Lys Gly Glu Phe Arg Leu Thr Gly ser Gly Phe Pro Ala Asp Gly pro 115 120 125 val Met Thr Asn ser Leu Thr Ala Ala Asp Trp cys val Asp Lys Leu 130 135 140

Leu Tyr Pro Asn Glu Asn Thr ile lie Gly Lys Phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr ser Gly Lys Arg Tyr Gin Ser Asp val Gin Thr Asn val 165 170 175

Thr Phe Gly Lys Pro lie ser Ala Asp lie Leu Lys Lys Gin Pro Met 180 185 190

Phe Val phe Arg Lys Val Glu Leu Lys His Ser Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp lie Val 210 215 220

<210> 84 <211 > 197 <212> PRT <213> Branchiostoma lanceolatum <400> 84

Met ser Leu Pro Ala Thr His Glu Leu His lie Phe Asp Lys lie Asn 15 10 15

Gly His Glu Phe Asp Met Arg Gly Lys Gly Thr Gly Asn Pro Asn Asp 20 25 30

Gly Tyr Glu Asp Leu Asp Leu Lys ser Thr Lys Asp Asp Leu pro phe 35 40 45 ser Pro Trp lie Leu val Gin Asn lie Gly Tyr Gly phe Asn Gin Tyr 50 55 60

Leu Pro Tyr Pro Asp Gly Ala Met ser Pro Phe Gin Ala Ala Met Tyr 65 70 75 80

Asn Gly Ser Gly Tyr His Val His Arg Glu Met Gly Phe Glu Asp Gly 85 90 95

Ala Thr Val Thr Gly lie Tyr Arg Tyr Thr Tyr Glu Gly ser His lie 100 105 110

Lys Gly Glu phe Gin val Asp Gly Thr Gly Phe Pro Ala Asp Gly Pro 115 120 125

Val Met Thr Asn Ser Leu Thr Asp Gin Asp Trp ser Val Thr Lys Met 130 135 140

Met Tyr Leu Asp Asn Lys Thr val Thr Ser Thr Ala Asp Gin Thr Tyr 145 150 155 160

Thr Thr Ala ser Gly Lys Arg Tyr Gin Gly Thr val Arg Thr Asn Asn 165 170 175

Thr phe Ala Lys Pro ile Ala Ala Asn ile Leu Gin Lys Gin Pro Val 180 185 190

Phe Val ser Arg Lys 195

<210> 85 <211> 196 <212> PRT <213> Branchiostoma lanceolatum <400> 85

Met ser Leu Pro Lys Thr His Glu Leu His ile Phe Gly Lys ile Asn 15 10 15

Gly His Glu Tyr Asp Met Arg Gly Lys Gly Thr Gly Asn pro Asn Asp 20 25 30

Gly Tyr Glu Asp Leu Asp Leu Lys ser Lys Asp Asp Leu Pro Phe ser 35 40 45

Pro Trp ile Leu val Gin Asn ile Gly Tyr Gly phe Asn Gin Tyr Leu 50 55 60

Pro Tyr Pro Asp Gly Ala Met Ser pro Phe Gin Ala Ala Met cys Asp 65 70 75 80

Gly ser Gly Tyr Glu val His Arg Glu Met Glu Phe Glu Asp Gly Ala 85 90 95

Thr Leu Thr Gly ile Tyr Arg Tyr Thr Tyr Glu Gly ser His ile Lys !00 105 110

Gly Glu Phe Gin val Asp Gly Thr Gly Phe Pro Asp Asp Gly Pro val -T5 120 125

Met Tan Ser Leu T^r AsP *-eu Asp Trp val val Thr Lys Met Val 130 135 140

Tyr pro Asp Glu Lys Thr val phe ser Thr ser Asp Gin Thr Tyr Thr 150 155 160

Thr Ala ser Gly Lys Gly Tyr Lys ser Thr val Arg Thr Asn Asn ile 165 170 175

Phe Ala Lys Pro Met Ala Ala Asp Met Met Gin Asn Gin Pro Ile Phe 180 185 190

Val ser Arg Lys 195

<210> 86 <211> 197 <212> PRT <213> Branchiostoma lanceolatum <400> 86

Met Ser Leu Pro Ala Thr His Glu Leu His ile Phe Gly Lys ile Asn 15 10 15

Gly His Glu Tyr Asp Met Arg Gly Lys Gly Thr Gly Asn Pro Asn Asp 20 25 30

Gly Tyr Glu Asp Leu Asp Leu Lys ser Thr Lys Asp Asp Leu Pro Phe 35 40 45 ser pro Trp ile Leu val Gin Asn ile Gly Tyr Gly phe Asn Gin Tyr 50 55 60

Leu Pro Tyr Pro Asp Gly Ala Met ser Pro Phe Gin Ala Ala Met cys 65 70 75 80

Asp Gly Ser Gly Tyr Glu Val His Arg Glu Met Glu Phe Glu Asp Gly 85 90 95

Ala Thr Val Thr Gly ile Tyr Arg Tyr Thr Tyr Glu Gly ser His ile 100 105 110

Lys Gly Glu phe Gin val Asp Gly Thr Gly Phe pro asp asp Giv Pro 115 120 125 val Met Thr Asp ser Leu Thr Asp Leu Asp Trp Val val Thr Lys Met 130 135 140 val Tyr Pro Asp Glu Lys Thr val Phe ser Thr ser Asp Gin Thr Tvr 145 150 155 160

Thr Thr Thr ser Gly Lys Gly Tyr Lys ser Thr val Arg Thr Asn Asn 165 170 175

Ile Phe Ala Lys Pro Ile Ala Ala Asp Met Met Gin ser Gin Pro val 180 185 190

Phe val ser Arg Lys 195

<210> 87 <211 > 197 <212> PRT <213> Branchiostoma lanceolatum <400> 87

Met ser Leu Pro Ala Thr His Glu Leu His Ile Phe Gly Lys lie Asn 15 10 15

Gly His Glu Phe Asp Met Arg Gly Lys Gly Thr Gly Asn Pro Asn Asp 20 25 30

Gly Tyr Glu Asp Leu Asp Leu Lys ser Thr Lys Asp Asp Leu Pro Phe 35 40 45 ser Pro Trp lie Leu val Gin Asn lie Gly Tyr Gly Phe Asn Gin Tyr 50 55 60

Leu Pro Tyr pro Asp Gly Ala Met ser Pro Phe Gin Ala Ala Met Tyr 65 70 75 80

Asn Gly ser Gly Tyr His val His Arg Glu Met Glu Phe Glu Asp Gly 85 90 95

Ala Thr Leu Thr Gly lie Tyr Arg Tyr Thr Tyr Glu Gly Ser His He 100 105 110

Lys Gly Glu Phe Gin Val Asp Gly Thr Gly Phe Pro Ala Asp Gly Pro 115 120 125 val Met Thr Asp ser Leu Thr Asp Leu Asp Trp Val val Thr Lys Met 130 135 140

Val Tyr Pro Asp Asp Lys Thr Val Phe ser Thr ser Asp Gin Thr Tyr 145 150 155 160

Thr Thr Thr Ser Gly Lys Gly Tyr Gin ser Thr val Arg Thr Asn Asn 165 170 175 lie Phe Ala Glu Pro He Ala Ala Asp Met Met Gin Ser Gin Pro val 180 185 190

Phe val ser Arg Lys 195

<210> 88 <211 >660 <212> DNA <213> Branchiostoma lanceolatum <220> <221 > mutation <222> (356)..(357) <223> AA instead of cc in wild-type sequence <400> 88 atgtctctcc cagcaaccca cgatttacac atctccggct caatcaatgg acatgagttt 60 gacttggaag gcagtggcaa gggcaatgca aaagaaggtt atcaggagct ccacctaaag 120 tccaacaagg gtgacetgtc attctccccc tggatcctgg tcccaaacat cggctacggc 180 ttctaccagt acctgccctt ccccgacgga gcgatgtcgc cttaccaggc cgccatgcac 240 gatggctccg gatacgtgat gcatcgttca atgcagtttg aggatggtgc catgctgcat 300 tcagaccacc gctacatcta taagggaaac catatcaaag gagagtttcg gctgaaagga 360 agcggtttcc ctgctgacgg ccctgtgatg accaactcgc tgaccgctgc ggactggtgc 420 gtcgacaagc tgctgtaccc aaacgacaac accataatcg gcaaattcga ctggacctac 480 accactacca gtggcaagcg ctaccaaagt gatgtgcaga ccaacgtcac atttggcaag 540 ccaatagcgg ccgacatttt gaagaagcag ccaatgttcg tgttccgcaa ggtggaactc 600. aagcacacca agactgagct caacttcaag cagtggcaga aggcattcca ggacatcgcc 660

<210> 89 <211> 660 <212> DNA <213> Branchiostoma lanceolatum <220> <221 > mutation <222> (356)..(357) <223> AA instead of cc in wild-type sequence <220> <221 > mutation <222> (524)..(524). <223> c instead of A in wild-type sequence <400> 89 atgtctctcc cagcaaccca cgatttacac atctccggct caatcaatgg acatgagttt 60 gacttggaag gcagtggcaa gggcaatgca aaagaaggtt atcaggagct ccacctaaag 120 tccaacaagg gtgacctgtc attctccccc tggatcctgg tcccaaacat cggctacggc 180 ttctaccagt acctgccctt ccccgacgga gcgatgtcgc cttaccaggc cgccatgcac 240 gatggctccg gatacgtgat gcatcgttca atgcagtttg aggatggtgc catgctgcat 300 tcagaccacc gctacatcta taagggaaac catatcaaag gagagtttcg gctgaaagga 360 agcggtttcc ctgctgacgg ccctgtgatg accaactcgc tgaccgctgc ggactggtgc 420 gtcgacaagc tgctgtaccc aaacgacaac accataatcg gcaaattcga ctggacctac 480 accactacca gtggcaagcg ctaccaaagt gatgtgcaga ccaccgtcac atttggcaag 540 ccaatagcgg ccgacatttt gaagaagcag ccaatgttcg tgttccgcaa ggtggaactc 600 aagcacacca agactgagct caacttcaag cagtggcaga aggcattcca ggacatcgcc 660

<210> 90 <211 >660 <212> DNA <213> Branchiostoma lanceolatum <220> <221 > mutation <222> (356)..(357) <223> AA instead of cc in wild-type sequence <220> <221 > mutation <222> (469)..(469) <223> A instead of G in wild-type sequence <220> <221 > mutation <222> (471)..(471) <223> G instead of C in wild-type sequence <220> <221 > mutation <222> (524)..(524) <223> C instead of A in wild-type sequence <400> 90 atgtctctcc cagcaaccca cgatttacac atctccggct caatcaatgg acatgagttt 60 gacttggaag gcagtggcaa gggcaatgca aaagaaggtt atcaggagct ccacctaaag 120 tccaacaagg gtgacctgtc attctccccc tggatcctgg tcccaaacat cggctacggc 180 ttctaccagt acctgccctt ccccgacgga gcgatgtcgc cttaccaggc cgccatgcac 240 gatggctccg gatacgtgat gcatcgttca atgcagtttg aggatggtgc catgctgcat 300 tcagaccacc gctacatcta taagggaaac catatcaaag gagagtttcg gctgaaagga 360 agcggtttcc ctgctgacgg ccctgtgatg accaactcgc tgaccgctgc ggactggtgc 420 gtcgacaagc tgctgraccc aaacgacaac accataatcg gcaaattcaa gtggacctac 480 accactacca gtggcaagcg ctaccaaagt gatgtgcaga ccaccgtcac atttggcaag 540 ccaatagcgg ccgacatttt gaagaagcag ccaatgttcg tgttccgcaa ggtggaactc 600 aagcacacca agactgagct caacttcaag cagtggcaga aggcattcca ggacatcgcc 660

<210> 91 <211> 220 <212> PRT <213> Branchiostoma lanceolatum <220> <221 > MUTAGEN <222> (119)..(119) <223> Lys instead of Thr in wild-type sequence <400> 91

Met ser Leu Pro Ala Thr His Asp Leu His- Ile ser Gly Ser lie Asn 1 5 10 15

Giv His Glu Phe Asp Leu Glu Gly ser Gly Lys Gly Asn Ala Lys Glu 20 25 30

Giv Tyr Gin Glu Leu His Leu Lys ser Asn Lys Gly Asp Leu ser Phe 35 40 45 ser Pro Trp Ile Leu val Pro Asn ile Gly Tyr Gly phe Tyr Gin Tyr 50 55 60

Leu Pro Phe Pro Asp Gly Ala Met ser Pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly Ser Gly Tyr val Met His Arg ser Met Gin Phe Glu Asp Gly 85 90 95

Ala Met Leu His ser Asp His Arg Tyr ile Tyr Lys Gly Asn His ile 100 105 110

Lys Gly Glu Phe Arg Leu Lys Gly ser Gly Phe Pro Ala Asp Gly Pro 115 120 125

Val Met Thr Asn Ser Leu Thr Ala Ala Asp Trp cys val Asp Lys Leu 130 135 140

Leu Tyr Pro Asn Asp Asn Thr Ile ile Gly Lys Phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr Ser Gly Lys Arg Tyr Gin ser Asp val Gin Thr Asn val 165 170 175

Thr Phe Gly Lys Pro ile Ala Ala Asp ile Leu Lys Lys Gin Pro Met 180 185 190

Phe Val Phe Arg Lys Val Glu Leu Lys His Thr Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp ile Ala 210 215 220

<210> 92 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <220> <221 > MUTAGEN <222> (119)..(119) <223> Lys in stead of Thr in wild-type sequence <220> <221 > MUTAGEN <222> (175)..(175) <223> Thr in stead of Asn in wild-type sequence <400> 92

Met Ser Leu Pro Ala Thr His Asp Leu His ile Ser Gly ser ile Asn 15 10 15

Gly His Glu phe Asp Leu clu Gly ser Gly Lys Gly Asn Ala lvs Glu 20 25 BO

Gly Tyr Gin Glu Leu His Leu Lys Ser Asn Lys Gly Asp Leu ser Phe 35 40 45 ser Pro Trp ile Leu val Pro Asn ile Gly Tyr Gly phe Tyr Gin Tyr 50 55 50

Leu Pro Phe pro Asp Gly Ala Met ser pro Tyr Gin Ala Ala Met His 65 70 75 80

Asp Gly Ser Gly Tyr Val Met His Arg ser Met Gin Phe Glu Asp Gly 85 90 95

Ala Met Leu His ser Asp His Arg Tyr ile Tyr Lys Gly Asn His ile 100 105 110

Lys Gly Glu Phe Arg Leu Lys Gly Ser Gly phe Pro Ala Asp Gly Pro 115 120 125

Val Met Thr Asn ser Leu Thr Ala Ala Asp Trp cys Val Asp Lys Leu 130 135 r 140

Leu Tyr Pro Asn Asp Asn Thr Ile ile Gly Lys Phe Asp Trp Thr Tyr 145 150 155 160

Thr Thr Thr Ser Gly Lys Arg Tyr Gin Ser Asp Val Gin Thr Thr Val 165 170 175

Thr Phe Gly Lys Pro Ile Ala Ala Asp ile Leu Lys Lys Gin Pro Met 180 185 190

Phe Val Phe Arg Lys Val Glu Leu Lys His Thr Lys Thr Glu Leu Asn 195 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp ile Ala 210 215 220

<210> 93 <211 >220 <212> PRT <213> Branchiostoma lanceolatum <220> <221 > MUTAGEN <222> (119)..(119) <223> Lys instead of Thr in wild-type sequence <220> <221 > MUTAGEN <222> (157)..(157) <223> Lys instead of Asp in wild-type sequence <220> <221 > MUTAGEN <222> (175)..(175) <223> Thr instead of Asn in wild-type sequence <400> 93

Met ser Leu Pro Ala Thr His Asp Leu His He Ser Gly Ser Ile Asn 15 10 15

Gly His Glu Phe Asp Leu Glu Gly Ser Gly Lys Gly Asn Ala Lys Glu M 25 30

Gly Tyr Gin Glu Leu His Leu Lys Ser Asn Lys Gly Asp Leu ser Phe 35 40 1 45r

Ser Pro Trp He Leu Val Pro Asn ile Gly Tyr cly phe Tyr Gin Tyr )U 55 60

Leu Pro Phe Pro Asp Gly Ala Met Ser pro Tyr Gln Ala Ala Met His 65 70 75 80

Asp Gly Ser Gly T^r Val Met His Arg Ser Met Gin Phe Glu Asp cly

Ala Met Leu His ser Asp His Arg Tyr lie Tyr Lys Gly Asn His Ile 100 105 * * no

Lys Gly Glu Phe Arg Leu Lys Gly ser Gly Phe pro Ala Asp Gly Pro •115 120 125

Val Met Thr Asn ser Leu Thr Ala Ala Asp Trp cys val Asp Lys Leu 130 135 140

Leu Tyr pro Asn Asp Asn Thr Ile lie Gly Lys Phe Lys Trp Thr Tyr 145 y 150 155 160

Thr Thr Thr ser Gly Lys Arg Tyr Gin ser Asp val Gin Thr Thr val ΙΠΓ Inr 165 170 175

Thr ohe cly Lys Pro lie Ala Ala Asp lie Leu Lys Lys Gin Pro Met

Tnr pne b j 18Q 185 190

Phe wai phe Arg Lys Val Glu Leu Lys His Thr Lys Thr Glu Leu Asn pne vai g5 200 205

Phe Lys Gin Trp Gin Lys Ala Phe Gin Asp lie Ala 210 215 220

Claims (7)

1. Fluorescent protein, wherein the protein is an isolated mutant or recombined nant protein having at least 80% amino acid sequence identity with the fluorescent protein having an amino acid sequence selected from the group consisting of SEQ ID NOs: 24-46, 68-87 and 91-93, and has the internal amino acid sequence Gly Tyr Xaa Xaa Xaa Gin Tyr Leu Pro Xaa Pro (SEQ ID NO: 47) 1 5 10 wherein Xaa at position 3 is Ala or Gly, Xaa at position 4 is Phe, His or Tyr, Xaa in position 5 is His, Tyr or Asn, and Xaa at position 10 is Phe or Tyr.

2. An isolated and purified structural gene encoding a fluorescent protein according to claim 1, comprising internal amino acid sequence Gly Tyr Xaa Xaa Xaa Gin Tyr Leu Pro Xaa Pro (SEQ ID NO: 47) 1 5 10 wherein Xaa at position 3 is Ala or Gly, Xaa at position 4 is Phe, His or Tyr, Xaa in position 5 is His, Tyr or Asn, and Xaa at position 10 is Phe or Tyr.

3. The structural gene of claim 2, wherein the structural gene is selected from the group consisting of SEQ ID NOs: 1 - 23rd

4. The structural gene according to claim 2, wherein the structural gene is selected from the group consisting of SEQ ID NOs: 48-67.

5. The structural gene of claim 2, wherein the structural gene is selected from the group consisting of SEQ ID NOs: 88-90.

6. A vector comprising a structural gene according to any one of claims 2-5.

7. A host cell comprising a vector according to claim 6 or a transgene containing a structural gene according to any one of claims 2-5.