JP2001333784A - Environmental stress-resistant gene - Google Patents

Abstract

PROBLEM TO BE SOLVED: To improve the resistance of various species of organisms, especially higher plants to environmental stresses. SOLUTION: A cDNA library derived from an organism having strong stress resistance such as mangrove is expressed in Escherichia coli having strong stress sensitivity to select a clone having enhanced stress resistance. A gene obtained by the method is expressed in various species of cells to thereby create a transformant cell having the enhanced stress resistance.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a DNA encoding a protein having an activity of improving tolerance to environmental stress such as salt stress, a method for screening the same, a protein having an activity of improving tolerance to environmental stress such as salt stress, and transgenic plants. And the use of these DNAs and proteins.

[0002]

2. Description of the Related Art Organisms existing in nature are exposed to various environmental stresses such as salt stress, high temperature stress, low temperature stress, freezing stress, and drought stress. In particular, salt stress is one of the major factors inhibiting the growth of many higher plants. Since enhancing the salt tolerance of higher plants leads to an increase in the production of agricultural products, attempts to enhance the salt tolerance of higher plants by gene transfer have been actively promoted in recent years.

[0003] For example, HJ Bohnert et al. Reported that the salt tolerance of tobacco was enhanced by introducing mannitol synthase derived from Escherichia coli into tobacco (Science VoI).
l.259, No.22, p508-510, 1993). Such enhanced salt tolerance of higher plants is based on proline synthase (Plant Physiol Vo
l.108, p1387-1394, 1995) and the introduction of a glycine betaine synthase enzyme has been shown to produce similar effects (Plant JVol. 12, p133-142, 1997, Plant).
Mol Biol Vol. 38, 1011-1019, 1998). However,
Recombinant plants obtained by introduction of genes encoding these enzymes are not sufficiently adaptable to salt water such as seawater.

[0004] Generally, environmental stress such as salt stress adversely affects a plurality of biological reactions. That is, in order to produce a genetically modified plant that can stably grow in an environment having a high salt concentration, it is necessary to use a group of genes encoding a plurality of proteins having activity to improve tolerance to salt stress. So far, the main method for isolating genes encoding proteins having an activity to improve environmental stress tolerance is based on the precondition that "the mechanism against stress is expressed when stress is applied". . Specifically, when environmental stress is exerted on a plant, a protein or mRNA that is specifically expressed is detected, a gene corresponding thereto is obtained by a molecular biological technique, and the gene is introduced into a plant that is vulnerable to stress. Therefore, a method of examining whether environmental stress tolerance can be seen has been used. Certainly, a gene that is specifically induced by environmental stress by this method has been isolated, but by introducing the gene into a plant that is vulnerable to environmental stress, a plant that is resistant to environmental stress can be produced. Cases are rare. For example, in a plant that grows under stress conditions with high salt concentration, if genes related to salt tolerance are expressed regardless of the presence or absence of stress, it is impossible to find stress tolerance genes by conventional methods. . In recent years, genome projects for plant species that are resistant to stress have been advanced, but once the nucleotide sequence or amino acid sequence is known, as in other genome projects, the functions are often unknown, and At present, it is not possible to specify whether or not is involved in environmental stress tolerance.

[0005] On the other hand, mangrove plants are trees that inhabit the soil containing high concentrations of salt near the coast and near the estuary.
Since mangrove plants are considered to have acquired a special salt tolerance mechanism during the evolution process, if the genes related to the salt tolerance of mangrove plants can be isolated, application to the enhancement of salt tolerance of higher plants can be expected. However, no examples of analyzing the salt tolerance mechanism of mangrove plants at the gene level have been known so far. The reason for this is that it was extremely difficult to extract mRNA of a gene directly involved in salt tolerance from such trees.

Recently, Mimura et al. Have established a cultured cell line of Bruguiera sexangula, a kind of mangrove plant (J.
Res Vol. 110, p25-29, 1997). These cultured cells have very specific properties different from other plant cultured cells, such as being capable of suspension culture and being able to grow stably even in an environment with a salt concentration of 150 mM or more (J . P
lant Res Vol. 110, p31-36, 1997). However, no attempt has been made to construct a cDNA library of a mangrove plant using the cultured cells to search for a group of genes related to salt tolerance of the mangrove plant. In addition, there have been almost no examples of enhancing the salt tolerance of higher plants by introducing genes derived from other halophytes, and ice plants (Ma
(sembryanthenum crystallinum), the inositol methyltransferase gene was introduced into tobacco to increase the content of ononitol, a kind of compatible solute, in the transformed cells, thereby enhancing the salt tolerance of tobacco (Plant Physiol Vol. .115, p1221-1219, 199
8) Alternatively, a gene encoding a barley-derived stress-inducing protein (LEA protein) having relatively high salt tolerance was introduced into rice to slightly improve the salt tolerance of rice (Plant Physiol Vol. 110). , p249-257, 1996) is a typical example. As described above, since a technology for efficiently isolating genes encoding proteins having salt stress tolerance improving activity has not been established, studies on environmental stress tolerance genes of many halophytes, including mangrove plants, have been conducted. It is hard to say that this has been done enough.

[0007] Further, if the function of a protein to improve its resistance to salt stress can be enhanced by artificially modifying a gene encoding a protein having an activity of improving its resistance to salt stress, a stronger salt stress resistance can be obtained. Plants can be created. When expressing choline dehydrogenase from E. coli in plants,
Glycinebetaine, a metabolite of choline dehydrogenase, is stabilized by modifying some codons to stabilize choline dehydrogenase expression in plants (a type of compatible solute that acts to increase plant salt tolerance) Attempts have been made to stabilize levels (Stress response
s of photosynthesis organisms (ed.Satoh K., Murat
a N.) p115-131, Elsevier Science, Amsterdam)
This is not a change in the amino acid sequence of the protein. There has been no report of a case in which the salt stress tolerance of a higher plant has been enhanced by introducing a protein having an activity of improving (improving) the salt stress, the amino acid sequence of which has been modified (improved). In addition, genes involved in salt tolerance and their modified genes are not only resistant to salt stress, but also to all other environmental stresses (heat, freezing, osmotic pressure, drying, ultraviolet rays), or to some of them. It can be expected to have the activity to improve.

[0008]

An object of the present invention is to provide a method for efficiently screening genes having an effect of enhancing resistance of various organisms to environmental stress, and an activity for improving environmental stress resistance obtained by such a screening method. It is an object of the present invention to provide a gene of a protein having an activity of improving environmental stress tolerance, a protein having an activity of enhancing environmental stress tolerance, and a transgenic plant having enhanced salt tolerance.

[0009]

Means for Solving the Problems In order to solve the above-mentioned problems, the present inventors focused on mangrove, a kind of halophyte, and found that a cultured cell line was already established in Bruguiera se
xangula cultures were obtained and this cell line was
The cells were cultured in the presence of mM NaCl, and a cDNA library was prepared based on mRNA extracted from the cultured cells, and a search was made for a gene involved in salt tolerance of mangroves from the library. Usually, a differential screening method is widely used as a method for screening for a salt tolerance-related gene (see Japanese Patent Application Laid-Open No. 10-295380 concerning a novel thionin gene induced by salt stress). Such a gene is a gene that is specifically induced under stress conditions, and expression of the gene in another cell cannot always enhance the stress tolerance of that cell. Therefore, the present inventors have newly developed a method of using a gene expression system of Escherichia coli for searching for a gene involved in stress tolerance.

When screening for genes involved in salt tolerance using the gene expression system of Escherichia coli, there is a problem that Escherichia coli itself has a strong defense mechanism against sodium chloride (NaCl). Currently, Escherichia coli such as DH5α, HB101, and JM109, which are widely used in the field of molecular biology, have a colony forming ability even on a 2YT agar medium containing 1000 mM or more of NaCl. When the above-mentioned screening was performed using these strains, in addition to the clones whose salt tolerance was enhanced by the expression of the candidate cDNA derived from the above cDNA library, the salt tolerance mechanism of Escherichia coli itself worked strongly and the salt tolerance was increased. As a result, a clone into which a cDNA which is not involved in any of the above is introduced will be obtained, and it is extremely difficult to clearly discriminate these two. For these reasons, selection of a salt tolerance-related gene using an Escherichia coli gene expression system has not been performed at all. The present inventors have found Escherichia coli having a reduced function of the salt tolerance mechanism as compared with other Escherichia coli, and have succeeded in screening a salt tolerance-related gene using Escherichia coli for the first time by using this.

The genes (cDNAs) derived from salty plants such as mangroves isolated by the above method by the present inventors are as follows:
It was confirmed that each had the function of enhancing the salt tolerance of Escherichia coli. Since the plant genes were expressed in Escherichia coli, which is a heterologous organism, and the salt tolerance of the Escherichia coli could be improved, these genes enhance salt tolerance in a wide range of organisms from prokaryotes to eukaryotes. It was considered to have a function. In fact, the present inventors introduced yeast, plant cells (tobacco cultured cells), and plant (tobacco cells) by introducing one of the isolated genes involved in stress tolerance, designated as the man1 gene. ) Has also succeeded in enhancing the salt resistance. In addition, it was also confirmed that manga1 has a function of enhancing resistance to environmental stresses such as heat, osmotic pressure, and freezing, in addition to salt resistance. Furthermore, a random mutation was introduced into the mang1 cDNA,
By introducing the mutant cDNA into Escherichia coli and repeating the step of selecting under stricter conditions than the selection conditions before the mutation one or more times, it has been found that a protein having stronger salt tolerance improving activity can be obtained. The present invention has been completed based on such a series of studies.

That is, the present invention relates to a method for screening a DNA encoding a protein having an activity for improving environmental stress tolerance, which comprises introducing a candidate cDNA derived from a cDNA library into a host cell and transforming the resulting transformed cell into a host cell. A screening method comprising culturing cells under environmental conditions in which cells cannot substantially grow, selecting clones growing after culturing, and isolating a candidate cDNA introduced from the selected clones (claim 1). Encoding a protein having activity to improve environmental stress tolerance
A method for screening A, comprising introducing a candidate cDNA from a cDNA library into a host cell, culturing the resulting transformed cell under environmental conditions in which the host cell cannot substantially grow, and growing after culturing. Cloned from the selected clone, isolating the introduced candidate cDNA from the selected clone, introducing a random mutation into the isolated candidate cDNA, introducing the mutated cDNA into a host cell,
One step is to select under more severe conditions than DNA selection conditions.
Or a screening method characterized by repeating twice or more times (claim 2), wherein the environmental stress is chemical stress, high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress, radiation stress, osmotic stress. The screening method according to claim 1 or 2, wherein the stress is one or two or more stresses selected from the group consisting of:
The screening method according to claim 3, wherein the stress is salt stress (claim 4), and the screening method according to any one of claims 1 to 4, wherein the host cell is Escherichia coli (claim 5). The screening method according to claim 5, wherein the Escherichia coli is a SOLR strain (claim 6), wherein the environmental condition under which host cells cannot substantially grow is a condition with a salt concentration of 350 mM or more. A screening method according to any one of claims 1 to 6 (claim 7).

[0013] The present invention also provides a DNA encoding a protein having an activity for improving environmental stress tolerance, which is obtained by the screening method according to any one of claims 1 to 7 (claim 8). Chemical stress, high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress,
1 or 2 selected from radiation stress and osmotic stress
9. A DNA encoding a protein having an activity for improving environmental stress tolerance according to claim 8, wherein the stress is the stress described above, and the chemical stress is a salt stress. Encoding a protein having the activity of improving environmental stress tolerance according to the above description
(Claim 10) The DNA encoding a protein having an environmental stress tolerance improving activity according to any one of claims 8 to 10, wherein the protein having an activity for improving environmental stress tolerance is a plant-derived protein. Claim 11), wherein the plant is a bilberry (Bruguiera sexangla),
Stag beetle (Avicennia marina), turkey (Suea)
DNA encoding a protein having environmental stress tolerance-enhancing activity according to claim 11, characterized in that the DNA is (a) or (d) a japonica), a swordfish (Salsola komarovii) or an ice plant (Mesembryanthemum crystallinum). (A) a protein comprising the amino acid sequence shown in SEQ ID NO: 2; (b) an amino acid having 70% or more homology with the amino acid sequence shown in SEQ ID NO: 2; A protein having a salt stress tolerance-enhancing activity, and (c) an amino acid sequence represented by SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted or added, and at least A protein having an activity of improving salt stress tolerance (claim 1);
3) a DN sequence comprising the nucleotide sequence shown in SEQ ID NO: 1 or a sequence complementary thereto or a part or all of these sequences
A (claim 14), a DNA that hybridizes with the DNA of claim 14 under stringent conditions and encodes a protein having at least a salt stress tolerance improving activity (claim 15); (b) a DNA encoding the protein of any one of (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 4, and (b) a deletion of one or several amino acids in the amino acid sequence shown in SEQ ID NO: 4 A protein comprising a substituted or added amino acid sequence and having at least a salt stress tolerance improving activity (claim 16), the base sequence shown in SEQ ID NO: 3 or its complementary sequence, or a part or all of these sequences Containing DNA (claim 17), hybridizing with the DNA of claim 17 under stringent conditions, and DNA encoding a protein having Kutomo salt stress tolerance enhancing activity (claim 18),
DNA encoding any one of the following proteins (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 6, (b) one or more amino acids in the amino acid sequence shown in SEQ ID NO: 6 Protein consisting of an amino acid sequence in which amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress resistance (claim 19), the base sequence shown in SEQ ID NO: 5 or a complementary sequence thereof, or a sequence thereof D containing part or all of
NA (claim 20), a DNA that hybridizes with a DNA constituting the gene according to claim 20 under stringent conditions, and that encodes a protein having at least a salt stress tolerance improving activity (claim 21);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 8, (b) an amino acid sequence shown in SEQ ID NO: 8 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 2
2), a nucleotide sequence shown in SEQ ID NO: 7 or a sequence complementary thereto, or a DN containing a part or all of these sequences
A (claim 23), a DNA that hybridizes with the DNA constituting the gene according to claim 23 under stringent conditions and encodes a protein having at least a salt stress tolerance improving activity (claim 24);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 10, (b) an amino acid sequence shown in SEQ ID NO: 10 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 2
5), the nucleotide sequence shown in SEQ ID NO: 9 or its complementary sequence, or a DN containing a part or all of these sequences
A (claim 26), a DNA that hybridizes with the DNA constituting the gene according to claim 26 under stringent conditions and encodes a protein having at least a salt stress tolerance-enhancing activity (claim 27);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 12, (b) an amino acid sequence shown in SEQ ID NO: 12 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 2
8), a base sequence represented by SEQ ID NO: 11 or a sequence complementary thereto, or D containing a part or all of these sequences
NA (claim 29), a DNA that hybridizes with the DNA constituting the gene of claim 29 under stringent conditions, and that encodes a protein having at least a salt stress tolerance improving activity (claim 30);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 14, (b) an amino acid sequence shown in SEQ ID NO: 14 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 3
1), the base sequence shown in SEQ ID NO: 13 or its complementary sequence, or a D sequence containing a part or all of these sequences
NA (claim 32), a DNA that hybridizes with the DNA constituting the gene according to claim 32 under stringent conditions and encodes a protein having at least a salt stress tolerance improving activity (claim 33);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 16, (b) an amino acid sequence shown in SEQ ID NO: 16 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 3
4), a nucleotide sequence represented by SEQ ID NO: 15 or a sequence complementary thereto, or D containing a part or all of these sequences
NA (claim 35), DNA that hybridizes with the DNA constituting the gene according to claim 35 under stringent conditions, and that encodes a protein having at least a salt stress tolerance improving activity (claim 36);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 18, (b) an amino acid sequence shown in SEQ ID NO: 18 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 3
7), a base sequence represented by SEQ ID NO: 17 or a sequence complementary thereto, or D containing a part or all of these sequences
NA (claim 38), a DNA that hybridizes with the DNA constituting the gene according to claim 38 under stringent conditions and encodes a protein having at least a salt stress tolerance improving activity (claim 39);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 20, (b) an amino acid sequence shown in SEQ ID NO: 20 consisting of an amino acid sequence in which one or several amino acids have been deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 4)
0), the base sequence shown in SEQ ID NO: 19 or its complementary sequence, or a D sequence containing a part or all of these sequences
NA (claim 41), a DNA that hybridizes with the DNA constituting the gene according to claim 41 under stringent conditions, and that encodes a protein having at least a salt stress tolerance improving activity (claim 42);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 22, (b) an amino acid sequence shown in SEQ ID NO: 22 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 4)
3), a nucleotide sequence represented by SEQ ID NO: 21 or a sequence complementary thereto or a D sequence containing a part or all of these sequences
NA (claim 44), a DNA that hybridizes with the DNA constituting the gene according to claim 44 under stringent conditions, and that encodes a protein having at least a salt stress tolerance improving activity (claim 45);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 24, (b) an amino acid sequence shown in SEQ ID NO: 24 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 4)
6), a nucleotide sequence represented by SEQ ID NO: 23 or a sequence complementary thereto, or D containing a part or all of these sequences
NA (claim 47), a DNA that hybridizes with the DNA constituting the gene according to claim 47 under stringent conditions and encodes a protein having at least a salt stress tolerance improving activity (claim 48);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 26, (b) an amino acid sequence shown in SEQ ID NO: 26, which has an amino acid sequence in which one or several amino acids have been deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 4)
9), a base sequence represented by SEQ ID NO: 25 or a sequence complementary thereto or a D sequence containing a part or all of these sequences
NA (claim 50), a DNA that hybridizes with the DNA constituting the gene according to claim 50 under stringent conditions, and that encodes a protein having at least a salt stress tolerance improving activity (claim 51);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 28, (b) an amino acid sequence shown in SEQ ID NO: 28 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 5
2), a base sequence represented by SEQ ID NO: 27 or a sequence complementary thereto or a D sequence containing a part or all of these sequences
NA (claim 53), a DNA that hybridizes with the DNA constituting the gene according to claim 53 under stringent conditions and encodes a protein having at least a salt stress tolerance improving activity (claim 54);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 30, (b) an amino acid sequence shown in SEQ ID NO: 30 consisting of an amino acid sequence in which one or several amino acids have been deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 5
5), a base sequence represented by SEQ ID NO: 29 or a sequence complementary thereto, or D containing a part or all of these sequences
NA (claim 56), a DNA that hybridizes with the DNA constituting the gene according to claim 56 under stringent conditions and encodes a protein having at least a salt stress tolerance improving activity (claim 57);
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 32, (b) an amino acid sequence shown in SEQ ID NO: 32 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 5
8), a base sequence represented by SEQ ID NO: 31 or a sequence complementary thereto or a D sequence containing a part or all of these sequences
NA (claim 59), a DNA that hybridizes with the DNA constituting the gene according to claim 59 under stringent conditions, and that encodes a protein having at least a salt stress tolerance improving activity (claim 60):
D encoding the protein according to any one of (a) and (b)
NA (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 34, (b) an amino acid sequence shown in SEQ ID NO: 34 in which one or several amino acids are deleted, substituted or added, and A protein having at least a salt stress tolerance improving activity (claim 6)
1), the nucleotide sequence shown in SEQ ID NO: 33 or its complementary sequence, or a D sequence containing a part or all of these sequences
NA (claim 62), a DNA that hybridizes with the DNA constituting the gene according to claim 62 under stringent conditions, and that encodes a protein having at least a salt stress tolerance improving activity (claim 63):
D encoding the protein according to any one of (a) and (b)
NA (a) SEQ ID NOs: 36, 38, 40, 42, 44, 4
6,48,50,52,54,56,58,60,62
Or a protein consisting of the amino acid sequence shown in 64,
(B) SEQ ID NOs: 36, 38, 40, 42, 44, 46,
48, 50, 52, 54, 56, 58, 60, 62 or 64 consisting of an amino acid sequence in which one or several amino acids have been deleted, substituted or added, and at least improved salt stress resistance A protein having an activity (claim 64), SEQ ID NOs: 35, 3
7, 39, 41, 43, 45, 47, 49, 51, 5
A DNA comprising the nucleotide sequence shown in 3, 55, 57, 59, 61 or 63 or a complementary sequence thereof, or a DNA comprising part or all of these sequences (claim 65); a DNA constituting the gene according to claim 65; The present invention relates to a DNA (Claim 66) which hybridizes under stringent conditions and encodes a protein having at least a salt stress tolerance improving activity.

According to the present invention, there is provided a method for improving environmental stress tolerance using the DNA according to any one of claims 8 to 66 (claim 67), wherein the environmental stress is chemical stress, high-temperature stress, low-temperature stress. Stress, freezing stress, drying stress, ozone stress, ultraviolet light stress,
1 or 2 selected from radiation stress and osmotic stress
68. The method for improving environmental stress tolerance according to claim 67, wherein the stress is the above stress (claim 68), and the chemical stress is salt stress, wherein the chemical stress is salt stress. Improvement method (Claim 6
9).

The present invention also relates to a protein comprising the amino acid sequence represented by SEQ ID NO: 2 (claim 70), a protein comprising an amino acid sequence having 70% or more homology with the amino acid sequence represented by SEQ ID NO: 2 and at least salt stress A protein having a tolerance improving activity (Claim 71), comprising an amino acid sequence in which one or several amino acids are deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 2, and having at least a salt stress tolerance improving activity; Protein (claim 72), a protein consisting of the amino acid sequence shown in SEQ ID NO: 4 (claim 73), and one or several amino acids are deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 4 A protein comprising an amino acid sequence and having at least a salt stress tolerance improving activity ( Claim 74), a protein comprising the amino acid sequence represented by SEQ ID NO: 6 (Claim 75), and a protein comprising the amino acid sequence represented by SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted or added. A protein having an activity of improving salt stress tolerance (Claim 76), a protein comprising the amino acid sequence represented by SEQ ID NO: 8 (Claim 77),
A protein consisting of an amino acid sequence in which one or several amino acids are deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 8 and having at least an activity of improving salt stress resistance (claim 78);
A protein consisting of the amino acid sequence represented by
9) In the amino acid sequence represented by SEQ ID NO: 10,
A protein consisting of an amino acid sequence in which one or several amino acids have been deleted, substituted or added, and having at least a salt stress tolerance improving activity (claim 80); a protein consisting of the amino acid sequence shown in SEQ ID NO: 12 (claim 81) a protein comprising an amino acid sequence in which one or several amino acids have been deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 12, and having at least a salt stress tolerance improving activity (claim 8)
2) a protein consisting of the amino acid sequence shown in SEQ ID NO: 14 (claim 83), one or several amino acids deleted in the amino acid sequence shown in SEQ ID NO: 14,
A protein comprising a substituted or added amino acid sequence and having at least a salt stress tolerance improving activity (claim 84); a protein comprising the amino acid sequence represented by SEQ ID NO: 16 (claim 85); an amino acid sequence represented by SEQ ID NO: 16 A protein comprising an amino acid sequence in which one or several amino acids have been deleted, substituted or added, and having at least a salt stress tolerance improving activity (claim 86); a protein comprising an amino acid sequence represented by SEQ ID NO: 18 (claim 86) 87) a protein consisting of an amino acid sequence in which one or several amino acids are deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 18 and having at least a salt stress tolerance improving activity (claim 88) From the amino acid sequence shown in SEQ ID NO: 20 Protein (claim 89) that,
A protein consisting of an amino acid sequence in which one or several amino acids are deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 20 and having at least a salt stress tolerance improving activity (claim 90); SEQ ID NO: 2
A protein comprising the amino acid sequence represented by SEQ ID NO: 2 (claim 91), an amino acid sequence represented by SEQ ID NO: 22 in which one or several amino acids are deleted, substituted or added, and at least salt stress resistance A protein having an improving activity (Claim 92),
A protein comprising the amino acid sequence represented by SEQ ID NO: 24 (claim 93), a protein comprising the amino acid sequence represented by SEQ ID NO: 24 in which one or several amino acids have been deleted, substituted or added, and at least a salt thereof; A protein having an activity of improving stress tolerance (claim 94), a protein consisting of the amino acid sequence shown in SEQ ID NO: 26 (claim 95), one or several amino acids deleted in the amino acid sequence shown in SEQ ID NO: 26, A protein consisting of a substituted or added amino acid sequence and having at least a salt stress tolerance improving activity (claim 96), a protein consisting of the amino acid sequence shown in SEQ ID NO: 28 (claim 97), an amino acid shown in SEQ ID NO: 28 In the sequence, one or several amino acids are deleted or substituted. Or a protein comprising an added amino acid sequence and having at least a salt stress tolerance improving activity (claim 98), a protein comprising the amino acid sequence represented by SEQ ID NO: 30 (claim 99), an amino acid represented by SEQ ID NO: 30 A protein comprising an amino acid sequence in which one or several amino acids have been deleted, substituted or added in the sequence, and having at least a salt stress tolerance improving activity (claim 100); a protein comprising an amino acid sequence represented by SEQ ID NO: 32 (Claim 10
1) In the amino acid sequence represented by SEQ ID NO: 32,
A protein comprising an amino acid sequence in which one or several amino acids have been deleted, substituted or added, and having at least a salt stress tolerance improving activity (claim 102); a protein comprising the amino acid sequence shown in SEQ ID NO: 34 (claim 103), a protein consisting of an amino acid sequence in which one or several amino acids have been deleted, substituted or added in the amino acid sequence shown in SEQ ID NO: 34, and having at least a salt stress tolerance improving activity (claim 104); Numbers 36, 38, 40, 42, 44, 4
6,48,50,52,54,56,58,60,62
Or a protein consisting of the amino acid sequence represented by SEQ ID NO: 64 (Claim 105), SEQ ID NOs: 36, 38, 40, 42,
44, 46, 48, 50, 52, 54, 56, 58, 6
In the amino acid sequence represented by 0, 62 or 64, 1
Alternatively, the present invention relates to a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance (Claim 106).

Further, the present invention provides an antibody which specifically binds to the protein according to any one of claims 70 to 72 (claim 1).
07), an antibody that specifically binds to the protein of any one of claims 73 to 104 (claim 108),
The present invention relates to an antibody that specifically binds to the protein of claim 05 or 106 (claim 109), and the antibody of any of claims 107 to 109 (claim 110), wherein the antibody is a monoclonal antibody.

Further, the present invention provides a vector (claim 111) comprising a DNA encoding a protein having an activity for improving environmental stress tolerance according to any one of claims 8 to 12, and a vector according to claims 13 to 15. Any DN
A vector comprising A (claim 112),
A vector (claim 113) comprising the DNA according to any one of claims 16 to 63 (claim 113);
A vector (Claim 114) comprising the DNA of any one of claims 6 to 14.

The present invention also provides a transformed cell obtained by introducing a vector according to any one of claims 111 to 114 into a host cell (claim 11).
5) The host cell is a plant cell, wherein the transformed cell according to claim 115 (claim 116), or the transformed cell according to claim 115 or 116 is cultured, and the transformed cell or the transformed cell is cultured. The present invention relates to a method for producing a protein having an activity for improving environmental stress tolerance, which comprises recovering a recombinant protein from a supernatant of a culture solution (claim 117).

The present invention also provides a method for introducing a DNA encoding a protein having an activity of improving environmental stress tolerance according to any one of claims 8 to 12 into a plant cell, and carrying out division / proliferation and regeneration of the plant cell. And a transgenic plant (claim 11).
8) A transgenic plant obtained by introducing the DNA according to any one of claims 13 to 15 into a plant cell, and causing the plant cell to divide, proliferate, and redifferentiate (claim 119). ), A transgenic plant obtained by introducing the DNA according to any one of claims 16 to 63 into a plant cell and allowing the plant cell to divide, proliferate, and redifferentiate (claim 120). ,
A transgenic plant (Claim 121), which is obtained by introducing the DNA according to any one of claims 64-66 into a plant cell and allowing the plant cell to divide, proliferate, and redifferentiate. A transgenic plant (Claim 122), which is obtained by introducing the vector according to any one of Items 111 to 114 into a plant cell, and causing the plant cell to divide, proliferate, and redifferentiate.
The environmental stress is one or more stress selected from chemical stress, high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress, radiation stress, and osmotic stress. The transgenic plant according to any one of claims 118 to 122 (Claim 123), and the chemical substance stress is a salt stress. The transgenic plant according to Claim 123 (Claim 124), Claims 118 to 122. A propagation material characterized by being derived from the transgenic plant according to any one of claims 1 to 4.
Regarding 5).

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION As a method for screening a DNA encoding a protein having an activity of improving environmental stress tolerance according to the present invention, a candidate cDNA derived from a cDNA library is used.
DNA is introduced into a host cell, the resulting transformed cell is cultured under environmental conditions in which the host cell cannot substantially grow, a clone growing after the culture is selected, and a candidate cDNA introduced from the selected clone is introduced. There is no particular limitation as long as it is a screening method for isolating. This screening method can also be used for enhancing the function of a protein having an activity for improving environmental stress tolerance obtained by this method. That is, as a method for screening a DNA encoding a protein having an activity of improving environmental stress tolerance according to another embodiment of the present invention, a candidate cDNA derived from a cDNA library is introduced into a host cell, and the obtained transformed cell is subjected to host cell Is cultured under environmental conditions that do not substantially grow, clones growing after culture are selected, candidate cDNAs introduced from the selected clones are isolated, and random mutations are introduced into the isolated candidate cDNAs. A screening method characterized by repeating the step of introducing this mutant cDNA into host cells and selecting it under severer conditions than the selection conditions of the cDNA before mutation one or more times.

[0021] By repeatedly producing a gene fragment obtained by introducing a random mutation into the gene fragment obtained as a result of the above-described primary screening, and selecting this gene under conditions more severe than the screening conditions, the environmental fragmentation is repeated. As a method for introducing a random mutation into any gene fragment in a method for improving the function of a protein having a stress tolerance improving activity, a method utilizing PCR is generally used. To reduce fidelity by adding (AJournal
of Methods in Cell and Molecular Biology Vol1, p1
1-15 (1989), Yeast Vol 8, p79-82 (1992)) is the simplest. As another random mutation method, DNA shuffling (Proc Natl Acad Sci USA, Vol.
91, p10747-10751, 1994) can also be used.

The environmental stress may be any stress as long as it is a stress based on environmental factors. Specifically, chemical stress, high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress, Radiation stress, osmotic stress and the like can be exemplified. These environmental stresses may be stress based on a single factor or stress based on a plurality of factors. Also, as chemical substance stress,
The stress is not particularly limited as long as the stress is caused by a chemical substance, and examples thereof include salt stress and toxic substance stress.

The cDNA library is not particularly limited as long as it contains cDNAs of genes involved in environmental stress, such as those derived from plants, animals, microorganisms and the like. For example, when screening for a DNA encoding a protein having an activity to improve salt stress tolerance, halophytes such as mangroves (Pomegranate, Ohirugi, Mehirugi, Yaeyamahirugi, Hirugimodoki, Hirugidamashi, Nipa palm), Matsubagiku,
A cDNA library prepared from a species such as turkey, euglena, hamcho, pine tree, or red clover can be used. As a method for preparing a cDNA library, any method may be used as long as it is known to those skilled in the art. For example, as described in the Examples, extraction of total mRNA from cells is performed by the method of Ostrem et al. (Plant Physics).
iol Vol. 84, p1270-1275, 1987), and poly (A) from the prepared mRNA.
+ RNA was purified by Oligotex-dT30 <su
per> (Daiichi Kagaku).
The cDNA library was purified poly (A) + R
Based on NA, ZAP-cDNA / Gigapack Cloning Kit (Str
(Atagene).

Examples of host cells into which candidate cDNAs to be screened derived from the above-mentioned cDNA library are introduced include microbial cells such as bacteria and yeasts, and animal and plant cells. Escherichia coli, Bacillus subtilis, Saccharomyces
Preferred are animal cells such as Serviche, BHK cells and the like, and among them, Escherichia coli, which is rich in such knowledge, is fast growing and easy to handle, is preferred. Also, as a host cell,
Transformed cells into which candidate cDNAs derived from the cDNA library have been introduced are preferably cells that cannot substantially grow under environmental conditions in which they can grow, such as salt-sensitive cells and thermosalt-sensitive cells. It can be prepared by screening or by mutating a wild strain.

Next, the transformed cells are transferred to the above host cells.
It is performed by introducing a candidate cDNA derived from a DNA library, and such an introduction method is not particularly limited as long as it is a known gene introduction method such as a transformation method and an electroporation method. Then, the transformed cells into which the candidate cDNA has been introduced are subjected to environmental conditions under which the host cells cannot substantially grow.
For example, the cells are cultured under high salt conditions, high temperature conditions, and dry conditions. Subsequently, clones growing after the culture are selected by a known method, and a candidate cDNA introduced from the selected clone can be isolated by a known method.

Hereinafter, the screening method of the present invention will be described more specifically with reference to the case where the environmental stress is salt stress and the host cell is Escherichia coli. As Escherichia coli used as a host cell, Escherichia coli having a reduced function of a salt-tolerant mechanism is preferable, and Escherichia coli having a minimum minimum growth inhibitory concentration against salt is more preferable. For example, 750 mM or more, preferably 500 mM or more, more preferably 350 mM or more.
It is preferable to use Escherichia coli whose colony formation is suppressed in a medium containing NaCl of mM or more, since the screening efficiency of the salt tolerance-related gene can be improved. It has been found by the present inventors that such NaCl-sensitive Escherichia coli can hardly grow on an agar medium containing a low concentration of salt (350 mM or more NaCl), a SOLR strain (TOYOBO, Stratagene), which is a kind of Escherichia coli.
And commercially available from Riken Gene Bank, Inc.). When using this SOLR strain,
The introduction of the candidate cDNA was performed using the ZAP-cDNA / Gigapack Clonin
g SOLR strain and ExA contained in Kit (Stratagene)
An in vivo excision system by ssist helper phage can be used, and all these operations can be easily performed by following the above kit manual.

The SOLR strain into which the candidate cDNA thus obtained has been introduced is cultured in a medium containing about 400 mM NaCl, and the grown cells are selected. Converted clones can be selected. For selection of such a clone, for example, culture may be performed at 37 ° C. for 8 to 20 hours on an agar medium or the like, and a colony formed on the agar medium may be selected. Isolation of cDNA from the selected clones is described, for example, in the literature (Current Protocols in Molecular Biology).
(Greene Publishing Associates and Wiley-Interscien
ce (1987)).
Can be performed by extracting

Screening for obtaining the desired transformed Escherichia coli can also be repeated several times. For example, Escherichia coli into which a cDNA library has been introduced is first cultured in a medium containing a salt with a minimum growth inhibitory concentration, and clones that grow under these conditions are selected (primary screening). Next, cDN was selected from the selected clones.
A is isolated, reintroduced into E. coli, and the E. coli is cultured in a medium containing a salt higher than the minimum growth inhibitory concentration, and clones that grow under these conditions are selected (secondary screening). By repeating screening in this way,
The efficiency of the isolation of the salt tolerance-related gene can be increased.
As described above, secondary screening can also be performed using a mutated cDNA in which a random mutation has been introduced into the isolated cDNA.

The DNA encoding the protein having the activity of improving environmental stress tolerance of the present invention is not particularly limited as long as it is a DNA obtained by the above screening method. For example, chemical stress such as salt stress, heat stress, etc. , Dry stress, ozone stress, ultraviolet stress, radiation stress, osmotic stress etc.
Alternatively, a DNA encoding a protein having an activity of improving resistance to two or more stresses can be mentioned. In particular, as the DNA encoding the protein having the activity of improving salt stress tolerance, DNA derived from plants, preferably DNA derived from salty plants such as oak (Brugfuiera sexangula) can be exemplified. Examples of the DNA encoding a protein having a salt stress tolerance improving activity derived from mangrove include SEQ ID NO: 1 in the sequence listing.
Specific examples include DNAs having the nucleotide sequence shown in 3, 5, 7, 9, 11, or 13, and also encode a protein derived from an ice plant (Mesembryanthemum crystallinum) and having a salt stress tolerance improving activity. As DNA, SEQ ID NOs: 15, 17, 19,
DNAs having the nucleotide sequences shown in Nos. 35 and 63 may be used as DNAs encoding a protein having a salt stress tolerance improving activity derived from turkey (Sueada japonica).
SEQ ID NOs: 21, 37, 39, 51, 53, 57 in the sequence listing
DNA having the nucleotide sequence shown in
sola komarovii) derived from a DNA having the nucleotide sequence shown in SEQ ID NO: 23, 25, 41, 47, 49, 59, 61 in the sequence listing. DNA encoding a protein having an activity to improve salt stress tolerance derived from a mangrove, Avicennia marina) includes SEQ ID NOs: 27, 29, 31, 33, 4 in the sequence listing.
DNA having the nucleotide sequence shown in 3,43,55,
Each can be specifically exemplified.

The DNA of the present invention includes a base sequence shown in SEQ ID NO: in the above sequence listing, a complementary sequence thereof, a DNA containing a part or all of these sequences,
A DNA that hybridizes with such DNA under stringent conditions and encodes a protein having at least a salt stress tolerance-enhancing activity, and SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, and 16 in the sequence listing. , 18,2
0,22,24,26,28,30,32,34,3
6,38,40,42,44,46,48,50,5
DNA encoding a protein consisting of the amino acid sequence shown in 2, 54, 56, 58, 60, 62 or 64
A DNA encoding a protein having an amino acid sequence having 70% or more homology with the amino acid sequence shown in SEQ ID NO: 2 of the sequence listing and having at least an activity of improving salt stress tolerance; , 6,8,1
0,12,14,16,18,20,22,24,2
6,28,30,32,34,36,38,40,4
2,44,46,48,50,52,54,56,5
A DN comprising an amino acid sequence represented by 8, 60, 62 or 64, wherein one or several amino acids are deleted, substituted or added, and which encodes a protein having at least a salt stress tolerance improving activity
A. There has been no report that salt tolerance and the like have been enhanced by introducing these DNAs into various organisms, and it has been found for the first time by the present inventors.

The proteins of the present invention include SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 1 in the sequence listing.
8, 20, 22, 24, 26, 28, 30, 32, 3
4,36,38,40,42,44,46,48,5
0, 52, 54, 56, 58, 60, 62 or 64, or an amino acid sequence having 70% or more homology with the amino acid sequence shown in SEQ ID NO: 2 in the sequence listing, and A protein having at least an activity of improving salt stress tolerance, and SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, and
0,22,24,26,28,30,32,34,3
6,38,40,42,44,46,48,50,5
A protein comprising an amino acid sequence shown in 2, 54, 56, 58, 60, 62 or 64 in which one or several amino acids have been deleted, substituted or added, and having at least an activity of improving tolerance to salt stress; Can be mentioned.

The above SEQ ID NOs: 8, 14, 36, 38, 4
0, 42, 44, 46, 48, 50, 52, 54, 5
The amino acid sequence shown in 6, 58, 60, 62 or 64 is not considered to encode the full length of the protein, however, since the amino acid sequence itself has an activity of enhancing salt stress tolerance, the function of each full length protein is considered. It can be said that it is an area. As described above, the present invention includes a full-length protein containing these functional regions and a DNA encoding the full-length protein. And the partial length cD
As a method for isolating full-length cDNA based on NA, for example, Marathon cDNA Amplification Kit (Clonte
ch), 3'-Full RACE Core Set (Takara Shuzo), 5'-Fu
It is appropriate to use a kit such as ll RACE Core Set (Takara Shuzo Co., Ltd.) and follow these instructions.

As described above, the present invention provides SEQ ID NO: 2,
4,6,8,10,12,14,16,18,20,2
2,24,26,28,30,32,34,36,3
8, 40, 42, 44, 46, 48, 50, 52, 5
DNAs encoding proteins functionally equivalent to the protein consisting of the amino acid sequence shown in 4, 56, 58, 60, 62 or 64 are included. In general, it is well known that in a protein having a physiological activity, even if a small number of amino acids have substitutions, deletions, or insertions, the physiological activity may be maintained in some cases. Various well-known methods are well known, and some kits are already commercially available. For example,
Synthesize the primer into which the mutation has been introduced and use QuikChange Site-
Directed Mutagenesis Kit (Stratagene)
Can be used to easily mutate amino acids in a protein.

As described above, SEQ ID NOs: 2, 4,
6, 8, 10, 12, 14, 16, 18, 20, 22,
24, 26, 28, 30, 32, 34, 36, 38, 4
0, 42, 44, 46, 48, 50, 52, 54, 5
6, 58, 60, 62 or 64, even if the protein has an amino acid sequence in which one or more amino acids are substituted, deleted, added and / or inserted, various types of biological cells (for example, As long as they have an activity of enhancing environmental stress tolerance including at least salt stress tolerance of plant cells, Escherichia coli, yeast and the like, they are all included in the scope of the present invention.

In addition, SEQ ID NOs: 2, 4, 6, 8, 10, 1
2,14,16,18,20,22,24,26,2
8, 30, 32, 34, 36, 38, 40, 42, 4
4,46,48,50,52,54,56,58,6
DNA encoding a protein functionally equivalent to the protein having the amino acid sequence of 0, 62 or 64
Uses hybridization technology and gene amplification technology (Mo
lecular Cloning, a Laboratory Manual, Second Editi
on, Cold Spring Harbor Laboratory Press (1989)). For example, SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 2
0,22,24,26,28,30,32,34,3
6,38,40,42,44,46,48,50,5
SEQ ID NOs: 1,3,5,7,9,11,13,15,17,19, which encode proteins consisting of the amino acid sequence represented by 2,54,56,58,60,62 or 64
21,23,25,27,29,31,33,35,3
7, 39, 41, 43, 45, 47, 49, 51, 5
Using all or a part of the nucleotide sequence shown in 3, 55, 57, 59, 61 or 63 as a probe, hybridization is carried out under stringent conditions on DNA libraries derived from various organisms, and the probe is hybridized. By isolating the soybean DNA, it is possible to obtain a DNA encoding a protein having the desired activity of improving environmental stress tolerance including at least salt stress tolerance.

Hybridization conditions for obtaining such DNA include, for example, hybridization at 42 ° C. and washing at 42 ° C. with a washing buffer containing 1 × SSC and 0.1% SDS. Hybridization at 65 ° C. and washing at 65 ° C. with a washing buffer containing 0.1 × SSC and 0.1% SDS can be more preferably mentioned. The factors that affect the stringency of hybridization include various factors other than the above-mentioned temperature conditions, and those skilled in the art can appropriately combine various components and have the same properties as the above-described stringency of hybridization. Stringency can be realized.

In addition, SEQ ID NOs: 1, 3, 5, 7, 9, 1
1,13,15,17,19,21,23,25,2
7, 29, 31, 33, 35, 37, 39, 41, 4
3,45,47,49,51,53,55,57,5
At least the desired salt stress resistance can also be obtained by performing a polymerase chain reaction using DNA (or RNA) derived from various organisms as a template using an oligonucleotide prepared based on the information of the base sequence shown in 9, 61 or 63 as a primer. A DNA encoding a protein having an activity of improving environmental stress tolerance, including DNA, can be obtained.
In the present invention, such a hybridization technique or gene amplification technique is used as long as it has an activity of enhancing environmental stress resistance including at least salt stress resistance of various biological cells (eg, plant cells, Escherichia coli, yeast, etc.). And DNA that can be isolated.

The above SEQ ID NOs: 2, 4, 6, 8, 10, 1
2,14,16,18,20,22,24,26,2
8, 30, 32, 34, 36, 38, 40, 42, 4
4,46,48,50,52,54,56,58,6
A protein functionally equivalent to the protein consisting of the amino acid sequence represented by 0, 62 or 64 is considered to have high homology with the amino acid sequence represented by the above SEQ ID NO. High homology is 70%
Or more, preferably 80% or more, more preferably 90%
The above (for example, 95% or more) sequence identity. As described above, the present invention has a high homology in the amino acid sequence shown in the above SEQ ID NO: DNAs encoding proteins having an activity of enhancing stress tolerance are included. For example, in the amino acid sequence of SEQ ID NO: 2, since a region containing amino acids 1 to 86 has an activity of enhancing salt stress resistance, for example, all gene DNAs encoding an amino acid sequence containing this region fall within the scope of the present invention. included. The sequence homology can be determined, for example, by using the multi-alignment function of the genetic information processing software GENETYX-MAC (Software Development Co., Ltd.).

The method for improving the resistance to environmental stress according to the present invention includes the above-mentioned DNA of the present invention, ie, chemical stress such as salt stress, high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress, radiation stress. The method is not particularly limited as long as it uses a DNA encoding a protein having an activity of improving resistance to one or more environmental stresses selected from osmotic stress. Plants and animals and their tissues,
Environmental stress resistance of organs, cells, and microorganisms such as bacteria, yeast, and mold can be improved.

The chemical stress such as salt stress of the present invention, heat stress, drying stress, ozone stress, ultraviolet stress, radiation stress, osmotic stress, etc.
Alternatively, the antibody that specifically binds to a protein having two or more environmental stress tolerance improving activities may be any antibody that specifically binds to the protein of the present invention, and such an antibody may be a monoclonal antibody. Specific examples include immunospecific antibodies such as antibodies, polyclonal antibodies, chimeric antibodies, single-chain antibodies, humanized antibodies, and the like. SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 1
6,18,20,22,24,26,28,30,3
2,34,36,38,40,42,44,46,4
8, 50, 52, 54, 56, 58, 60, 62 or 6
4, a protein consisting of an amino acid sequence having 70% or more homology with the amino acid sequence shown in SEQ ID NO: 2 and having at least a salt stress tolerance improving activity, 6,
8, 10, 12, 14, 16, 18, 20, 22, 2
4,26,28,30,32,34,36,38,4
0, 42, 44, 46, 48, 50, 52, 54, 5
In the amino acid sequence shown in 6, 58, 60, 62 or 64, a protein comprising an amino acid sequence in which one or several amino acids have been deleted, substituted or added, and having at least a salt stress tolerance improving activity is used as an antigen. Can be manufactured. These antibodies are useful, for example, in elucidating the molecular mechanism of a protein having an activity to improve environmental stress tolerance.

Antibodies against the above-mentioned protein having an activity of improving environmental stress tolerance can be obtained by using a conventional protocol in an animal (preferably a non-human animal) to a fragment, analog or cell containing the protein or epitope having the activity of improving environmental stress tolerance. , For example, for the preparation of monoclonal antibodies, hybridoma technology (Nature 256, 495-497, 1975), trioma technology, human B-cell hybridomas, resulting in antibodies produced by continuous cell line cultures. Technique (Immunology Tod
ay 4, 72, 1983) and the EBV-hybridoma technique (MO
NOCLONAL ANTIBODIES AND CANCER THERAPY, pp.77-96,
Any technique such as Alan R. Liss, Inc., 1985) can be used.

In order to produce a single-chain antibody against the protein having the activity of improving environmental stress tolerance of the present invention,
The method for preparing single-chain antibodies (US Pat. No. 4,946,778) can be applied. Further, in order to express a humanized antibody, a transgenic plant or a transgenic animal or the like is used, or a clone that expresses a protein having the activity to improve its environmental stress tolerance is isolated and identified using the above antibody. The polypeptide can also be purified by affinity chromatography.

The vector of the present invention has the above-mentioned activity for improving tolerance to environmental stress such as chemical stress such as salt stress, high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress, radiation stress and osmotic stress. The vector is not particularly limited as long as it contains a DNA encoding a protein having the vector. The transformed cell of the present invention is not particularly limited as long as it can be obtained by introducing such a vector into a host cell such as a plant cell. Further, as a method for producing a protein having an activity of improving environmental stress tolerance according to the present invention, a method for culturing the above-mentioned transformed cell of the present invention and recovering a recombinant protein from the transformed cell or a supernatant of a culture solution thereof If so, there is no particular limitation. Furthermore, the transgenic plant of the present invention can be obtained by introducing a DNA encoding the protein having the activity for improving environmental stress tolerance or the vector into a plant cell, and allowing the plant cell to divide, proliferate, and redifferentiate. It is not particularly limited as long as it can be performed. Hereinafter, the above-described vector of the present invention, a transformed cell, a method for producing a protein having an activity to improve environmental stress tolerance, and a transgenic plant will be described.

As described above, the DNA of the present invention can be used for preparing a recombinant protein. To prepare a recombinant protein, the DNA of the present invention is inserted into an appropriate expression vector, and the vector is introduced into an appropriate host cell.
The expression can be carried out by expressing the DNA, and then recovering the expressed protein from the transformed cell or its culture supernatant. Host-vector systems used for expression of recombinant proteins include, for example, IMPACT-CN Sy
stem (host: E. coli strain ER2566, vector: pTYB)
1, pYB2, pYB11, pYB12 (BioLabs)) or pET Expression System (host: Epicurian Coli BL2)
1, vector: pET3 series (Novagen)). Methods for introducing a vector into a host cell include methods known to those skilled in the art, for example, an electroporation method and a heat shock method. , Academic Publishing Center (1998)). The culturing of the transformant for expressing the recombinant protein can be performed by a method and conditions generally used by those skilled in the art. The expressed protein is, for example, IMPACT-CN Sy
When using a stem, chitin beads (BioLabs
If the pET Expression System is used,
It can be purified by Bind Resin (Novagen).

The DNA of the present invention can also be used for producing a transgenic organism having at least enhanced salt stress tolerance. The species for producing the transgenic organism using the DNA of the present invention is not particularly limited, but when the gene to be used is derived from mangroves, it is preferably a higher plant. Such transgenic plants can be produced by inserting the DNA into a vector that guarantees its expression in plant cells, introducing the DNA into plant cells, and regenerating the transformed plant cells to obtain transgenic plants. It can be performed advantageously.

As a vector used for producing a transgenic plant, for example, pBI101 or pIG121Hm (Plant J, Vol 6, p271-2) commercially available from Toyobo.
82 (1994)). There is no particular limitation on the type of plant cell into which the vector is introduced, and for example, rice, wheat, corn, soybean, tobacco, carrot and the like can be considered. Examples of the form of plant cells include protoplasts, calli, and plant parts (leaf discs, hypocotyls, etc.). As a method for introducing a vector into a host plant cell, the Agrobacterium method is suitable, but in addition, for example, a polyethylene glycol method, an electroporation method, a particle gun method, or the like can be used (experiment with a model plant). Protocol, Shujunsha (1996)).

The method of regenerating a plant cell into which a vector has been introduced into a plant varies depending on the plant species. For example, in the case of rice, it can be performed as follows. Calli are induced from the ripe seeds, and the callus is infected with Agrobacterium into which the cDNA has been introduced. After co-cultivation, transfer to a selection medium and culture. After about 3 weeks, the calli are transferred to a regeneration medium and cultured until regeneration. After adaptation for 4 or 5 days, the transformant is regenerated by transferring to a pot (experimental protocol for model plants, Shujunsha (1996)). In addition, carrots, cigarettes, etc. can be regenerated by Kato,
Method of Dr. Shono et al. (Technology of plant tissue culture Asakura Shoten (198
3)) is appropriate.

The propagation material derived from the transgenic plant of the present invention may be any propagation material derived from the above-described transgenic plant. For example, seeds, tuberous roots, Examples include culture propagation materials such as cutting ears and mericlone. Also,
It is possible to mass-produce the transgenic plant of the present invention based on such a propagation material.

[0049]

EXAMPLES Hereinafter, the present invention will be described specifically with reference to examples, but the present invention is not limited to the following examples. Example 1 cDN of mangroves and other halophytes
Preparation of A library Mangrove suspension culture cells were obtained from Brug established by Mimura et al.
uiera sexangula suspension culture cell line (J Plant Res Vol.1
10, p25-29 (1997)). This cultured cell is 100
Using an AA medium containing mM NaCl, reciprocal shaking culture (70 rpm) was performed in a dark place at 26 ° C. in a volume of 120 ml in a 500 ml flask. A mangrove cDNA library was prepared by using the suspension cultured cells according to the following procedure. First, the method of Osttrem et al. (Plant Physiol Vol.
84 p1270-1275 (1987)), and
From here, use Oligotex-dT30 <super> (Daiichi Kagaku)
The oligo (A) + RNA was purified. Purified poly
(A) cDNA is synthesized based on + RNA, and λZapII
(Stratagene) Introduced into lambda phage vector and c
A DNA library was constructed. c using λZapII
The method for constructing the DNA library is a well-known method, and the actual procedure was in accordance with the manual of Stratagene. Mangrove cDNA containing the results, 10 ⁶ independent clones
Successfully constructed the library. Another halophyte, Hiragidashi (Avicenni, a type of mangrove)
a marina), turkey (Sueada japonica), swordfish (Salsolakomarovii), ice plant (Mesembryant
The leaves of each plant were used as a material for preparing a cDNA library of C. hemum crystallinum). cD
As a method for preparing the NA library, the same method as that in the above mangrove was used. Obtained cDNA
Library contained 10 ⁵ to 10 ⁶ independent clones, respectively.

Example 2 Determination of Screening Conditions for cDNAs Involved in Salt Tolerance As a method for screening cDNAs involved in salt tolerance from cDNA libraries of mangroves and other halophytes, the present inventors used E. coli gene expression. The system was used. That is, a novel method for obtaining cDNA related to salt tolerance was developed by introducing mangrove and other halophyte cDNAs into E. coli and selecting transformed Escherichia coli with enhanced salt tolerance. For the selection of transformed Escherichia coli with enhanced salt tolerance, 2
YT agar medium was used. When starting this screening, the present inventors aimed at selecting host Escherichia coli suitable for the above-mentioned screening and carried out various Escherichia coli (DH5
α, JM109, HB101, SOLR) was determined as the minimum growth inhibitory concentration for NaCl. These Escherichia coli are well-known strains, and are commercially available from TOYOBO, Stratagene, and the like. DH5α, JM109, and HB
The growth of 101 is markedly inhibited on a 2YT agar medium containing 1200 mM NaCl, but colony formation is possible, and the growth is completely suppressed by 1500 mM NaCl. In contrast, SOLR is 30
The growth was significantly suppressed by 0 mM NaCl, and 400 mM
NaCl completely inhibits its growth. From this, it was clarified that SOLR is a salt-sensitive strain and does not have a strong salt tolerance mechanism, unlike other Escherichia coli. The fact that the salt tolerance mechanism of the host Escherichia coli itself does not work strongly is very effective in performing the above screening. Therefore, screening of a cDNA library involved in salt tolerance from a cDNA library is carried out using S. coli as a host E. coli.
Using OLR, the NaCl concentration of the selective agar
M was set, and screening was performed according to the following procedure.

Example 3 cDNAs involved in salt tolerance from mangrove and other halophyte cDNA libraries
Screening of NA A cDNA library of mangroves and other halophytes was introduced into SOLR in a form incorporated into pBluescript SK by an in vivo excision system (Stratagene). For gene transfer, use ZAP-cDNA / Gigapack Cloning Kit
(Stratagene). In the present invention, the following two-step screening was performed in order to select the SOLR into which the cDNA related to salt tolerance was introduced from the SOLR into which cDNAs of mangrove and other halophytes were introduced. The primary screening was performed using SOLR introduced with mangrove and other halophyte cDNAs at 400 mM NaCl, 50 μg / ml.
ml kanamycin, 50 μg / ml ampicillin,
The cells were inoculated on a 2YT agar medium containing 0.05 mM IPTG and cultured at 37 ° C. for 20 hours. All the colonies obtained under these conditions were again inoculated on the agar medium, and their growth was observed. As a result, in Bruguiera, 168 clones with enhanced salt tolerance were obtained. Approximately the same number of clones with enhanced salt tolerance were successfully obtained in transformed Escherichia coli into which cDNA libraries of other halophytes were introduced. Since it was considered that some of these clones had enhanced salt tolerance mechanism derived from the host E. coli for some reason, the following secondary screening was performed.

Plasmids were extracted from each clone obtained in the primary screening, and all were reintroduced into SOLR. The newly obtained transformant contained 50 μg / ml kanamycin, 50 μg / ml ampicillin, 0.05
The cells were cultured in a 2YT liquid medium containing mM IPTG until the logarithmic growth phase was reached, and a series diluted with the 2YT liquid medium was prepared and spotted on the agar selection medium at 25 μl each. After air-drying until the liquid was dried, the cells were cultured at 37 ° C. overnight. As a result, in the Bruguiera cDNA library, a clear improvement in salt tolerance was confirmed in 30 clones. As a representative example, FIG. 1 shows the results of a spot experiment on E. coli in which the cDNA shown in SEQ ID NO: 1 was introduced. Next, the nucleotide sequence of the cDNA introduced into these 30 clones was analyzed using the Thermo Sequenase Cycle Sequencing Kit (Amersham).
And DNA sequencer LIC-4000L (L
I-COR) and determined according to the manufacturer's instructions. As a result, Brugeier obtained from 30 clones
The acDNA was classified into seven types. That is, 23 cDNAs shown in SEQ ID NO: 1 and cDNA shown in SEQ ID NO: 3
, Two cDNAs shown in SEQ ID NO: 5, two cDNAs shown in SEQ ID NO: 7, and a cDN shown in SEQ ID NO: 9.
A, one cDNA shown in SEQ ID NO: 11 and one cDNA shown in SEQ ID NO: 13 were obtained.

Similarly, the cDNAs shown in SEQ ID NOs: 15, 17, 19, 35 and 63 were obtained from the ice plant.
From SEQ ID NOS: 21, 37, 39, 5
1,53,57, the cDNA shown in SEQ ID NOS: 23,25,41,47,49,59,61 from Okahijiki, and the cDNA shown in SEQ ID NO: 27,
CDNA shown in 29, 31, 33, 43, 45, 55
, Respectively.

Using the BLAST homology search program,
The homology search of the amino acid sequences encoded by these DNAs was performed. As a result, the amino acid sequence represented by SEQ ID NO: 2 was considered to be a novel protein because no homologous protein was registered in databases such as Swiss protein and PIR. Then, the present inventors named the novel protein (total amino acid number: 141) encoded by this cDNA mangulin, and named this gene mang1. Next, an attempt was made to determine the functional region of mangulin. 16, 42, 65, 87, 10
A stop codon at amino acid 9,142;
A subclone was prepared by introducing methionine (introducing a stop codon immediately before) at amino acids 35 and 49, and this was introduced into SOLR. The above spot experiment was carried out. It was clarified that this was an area related to the enhancement of salt resistance (FIG. 2).

The amino acid sequence shown in SEQ ID NO: 4 is Arab
t-complex polypeptide 1 of idopsis thaliana (pir JN0
448) and about 90% homology. The amino acid sequence shown in SEQ ID NO: 6 is obtained from Metallinthion of Ricinus communis.
It had about 80% homology with ein-like protein TYPE 2 (EMBL L02306). The amino acid sequence shown in SEQ ID NO: 8 is
It had about 63% homology with Homo sapiens RubB-like DNA helicase (AB024301). The amino acid sequence shown in SEQ ID NO: 10 is Ribosomalprotein S of Rattus norvegicus.
29 (pir S30298) had about 45% homology. The amino acid sequence shown in SEQ ID NO: 12 is Elongation f of Zea mays.
It had about 90% homology with actor eEF-1 alpha chain (pir S66339). The amino acid sequence shown in SEQ ID NO: 14 had about 70% homology with cdz21 (pir S26640) of Schizosaccharomyces pombe. SEQ ID NOs: 2, 4, 6, 8,
The cDNA shown in SEQ ID NO: 1, 3, 5, 7, 9, 11, or 13 encoding the protein shown in 10, 12, or 14, respectively, has a function of actually improving the salt tolerance of E. coli, It is thought to have the function of enhancing salt tolerance in a wide range of organisms including prokaryotes to higher plants.

Similarly, the amino acid sequence derived from the ice plant shown in SEQ ID NO: 16 is Arabidopsis thaliana
F13O11.15 gene product (gp AC006193_15) and 68%
With the homology of The amino acid sequence derived from the ice plant shown in SEQ ID NO: 18 is H + -t of Arabidopsis thaliana.
ransporting ATPase (EC 3.6.1.35) 14K chain (pir T0
1087) and 78% homology. The amino acid sequence derived from the ice plant shown in SEQ ID NO: 20 is Arabidopsis
thaliana 40S RIBOSOMAL PROTEIN S20 (pir T12992)
And 91% homology. The amino acid sequence derived from turpentine shown in SEQ ID NO: 22 is obtained from Atriplex canescens.
It had 63% homology with ozone-inducible protein (prf 2316438B). The amino acid sequence derived from Okamijiki shown in SEQ ID NO: 24 is GIBBERELLI of Arabidopsis thaliana.
It had 58% homology with N-REGULATED PROTEIN 1 PRECURSOR (sp GAS1_ARATH). The amino acid sequence derived from okajiki shown in SEQ ID NO: 26 is obtained from ADP-r of Vigna unguiculata.
It had 99% homology with ibosylation factor (gp AF022389_1). The amino acid sequence derived from Hiragidamashi shown in SEQ ID NO: 28 is a tuberization-ind of Solanum demissum.
It had 56% homology with uced protein (prf 2310431A). The amino acid sequence derived from Hiragidamashi shown in SEQ ID NO: 30 is the Enod93 protein of Medicago sativa (gp MSA248
334_1) and 69% homology. The amino acid sequence derived from Hiragidamashi shown in SEQ ID NO: 32 is Oryza sativa
40S RIBOSOMAL PROTEIN S21 (pir S38357) and 69%
With the homology of The amino acid sequence derived from Hiragidamashi shown in SEQ ID NO: 34 is Mesembryanthemum crystallinu
m protein phosphotase 2C homolog (AF097667) and 7
It had 9% homology. Mesembryanthemum crystallinu
m protein phosphotase 2C homolog (AF097667) and 79
% Homology.

The amino acid sequence derived from the ice plant shown in SEQ ID NO: 36 is the pRIB5 protein of Ribes nigrum.
n (gp RNI7578_1) with 58% homology. The amino acid sequence derived from radish as shown in SEQ ID NO: 38 is Me
sembryanthemum crystallinum tonoplast intrinsic p
It had 84% homology with rotein (pir T12439). The amino acid sequence derived from radish as shown in SEQ ID NO: 40 is
Spinacia oleracea phosphoethanolamine N-methyltra
It had 86% homology with nsferase (gp AF237633_1). The amino acid sequence derived from Okahijiki shown in SEQ ID NO: 42 is a phosphoenolpyruvate car of Selenicereus wittii.
It had 83% homology with boxylase (gpu SWI17843_1). The amino acid sequence derived from Hiragidamashi shown in SEQ ID NO: 44 is a putative chaperonin of Arabidopsis thaliana.
(Gp ATAC021640_16) with 84% homology. The amino acid sequence derived from Hirugidamushi shown in SEQ ID NO: 46 is
Arabidopsis thaliana hypothetical protein T5F17.4
0 (pir T10653) and 88% homology.

In addition, the amino acid sequence derived from Okahiziki shown in SEQ ID NO: 48 is composed of cysteine proteina of Glycine max.
It had 63% homology with se inhibitor (pir T07139). The amino acid sequence derived from Okahijiki shown in SEQ ID NO: 50 is a nucleotide sugar epime of Arabidopsis thaliana.
It had 87% homology with rase-like protein (gp ATCHRIV73_7). The amino acid sequence derived from turkey, shown in SEQ ID NO: 52, is a putative prototype of Arabidopsis thaliana.
It had 57% homology with tein (gp ATT20K12 — 12). The amino acid sequence derived from turkey, shown in SEQ ID NO: 54, is a putative WD-40 repeat protein of Arabidopsisthaliana.
in (gp AC006569_14) with 78% homology. The amino acid sequence derived from Hirugidamushi shown in SEQ ID NO: 56 is
Medicago sativa cdc2MsE gene product (gp MSCDC2MS
E_1) and 75% homology. The amino acid sequence derived from radish as shown in SEQ ID NO: 58 is Arabidopsis th
Aliana putative protein (gp ATF17C15_9) and 39%
With the homology of The amino acid sequence derived from Okahijiki shown in SEQ ID NO: 60 is transcripti of Nicotiana tabacum.
It had 66% homology with on factor E2F (prf 2601241A). The amino acid sequence derived from Okahijiki shown in SEQ ID NO: 62 is a hypothetical protein of Arabidopsis thaliana.
It had 34% homology with T26B15.5 (pir T02548).
The amino acid sequence derived from the ice plant shown in SEQ ID NO: 64 is Homo sapiens of Homo sapiens cDNA FLJ10298 fis,
clone NT2RM1001115, weakly similar to ENDOCHITINA
SE2 PRECURSOR (EC 3.2.1.14) (pir T02548) and 30%
With the homology of

Example 4 Mangrove cD in yeast
Effect of NA pBlu from which cDNA shown in SEQ ID NO: 1 was cloned
escript SK was digested with restriction enzymes EcoRI and NotI, and subjected to agarose gel electrophoresis. The fragment of about 1 kb obtained here was cut out and purified with GENECLEAN Kit (BIO101). This was introduced into a yeast expression vector pYES2 (Invitrogen) cut with restriction enzymes EcoRI and NotI using Ligation Kit ver2 (Takara Shuzo). Next, this was introduced into yeast by electroporation. Yeast is Saccharomyces cereviciae YM4271 (Clontec
h company) was used. For selection of transformed yeast, SD agar medium without uracil (hereinafter, -UraSD agar medium)
Was used. The salt tolerance of the transformed yeast was evaluated as follows. Transformed yeast cultured in -UraSD medium until late logarithmic growth phase contains -1200 mM NaCl -UrSD
Inoculation (initial concentration OD600 = 0.1) was performed on aSD medium and -UraSD medium without NaCl, and shaking culture was started at 30 ° C. The cell suspension was collected every 24 hours, and the absorbance was measured. The absorbance was measured using UV-1200 manufactured by Shimadzu Corporation. As a comparison, the same study was performed on yeast into which only the vector pYES2 was introduced. The result is shown in FIG. As is clear from FIG. 3, it was confirmed that the cDNA obtained as a result of the screening has a function of enhancing salt tolerance in yeast as in the case of Escherichia coli.

Example 5 Effect of Mangrove cDNA on Cultured Tobacco Cells pBluescriptSK, into which the cDNA shown in SEQ ID NO: 1 has been cloned, is replaced with restriction enzymes Xba1 and Xh.
This was cut with o1, and this was subjected to agarose gel electrophoresis. The fragment of about 1 kb obtained here was cut out and GENECLEAN Ki
Purified by t (BIO101). This is Ligation Kit ve
r2 (Takara Shuzo) using the plant expression vector pBI101
(EMBOJ Vol6, p3901-3907 (1987))
Introduced to otI site. The resulting plasmid was introduced into Agrobacterium by electroporation,
This was transformed into tobacco cultured cells (Nicotiana tabacum L. Cv. Bri
ght Yellow 2). A method for introducing a gene into a plant cell using Agrobacterium is a well-known method. Here, Agrobacterium tumefaci
ens EHA101 and the method of An (Plant Physiol Vol.7
9, p568-570 (1985)). The salt tolerance of the transformed tobacco cultured cells was evaluated as follows. The callus of the transformed cultured tobacco cells was picked up and cultured in Lins-Mayer medium until late logarithmic growth. This was adjusted to have NaCl concentrations of 0, 100, and 150 mM45.
The cells were subcultured at a rate of 1 ml each in ml Lins-Mayer medium and shaking culture was started at 26 ° C. Cells were collected from the cell suspension on day 7 or 13 after the start of culture, and the wet weight was measured. As a comparison, a similar study was carried out on cultured tobacco cells into which the GUS gene had been introduced using pBI101 instead of mangrove cDNA. FIG. 4 shows the results. As is clear from FIG. 4, it was confirmed that the cDNA obtained as a result of the screening has a function of enhancing salt tolerance in tobacco cultured cells as in the case of Escherichia coli.

Example 6 Effect of Mangrove cDNA on Tobacco (Plant) The plasmid obtained in Example 5 was introduced into Agrobacterium by electroporation, and this was used to infect a tobacco leaf disk. A method for introducing a gene into a tobacco leaf disk using Agrobacterium is a well-known method. Here, Agrobacterium
Using ium tumefaciens EHA101, the method described in Introduction to Plant Cell Engineering (Academic Publishing Center (1998)) was used. Evaluation of the salt tolerance of the transformed tobacco (plant) was performed as follows. Transformed tobacco was used at a NaCl concentration of 150 m.
M, and transferred to MS agar medium adjusted to M.
° C, under light irradiation of light-dark cycle (light: 16 hours / dark: 8 hours)
And cultured. The growth of the plant was observed 30 days after the start of the culture, and its salt tolerance was evaluated. Similar studies were performed on cultured tobacco cells into which the GUS gene had been introduced using pBI101 instead of mangrove cDNA. The results are shown in FIG. 5 (see reference pictures). As is clear from FIG. 5, the tobacco plant into which the cDNA obtained as a result of the screening was introduced exhibited roots and roots in the presence of NaCl.
The growth of leaves and stems was good. From this, it was confirmed that the cDNA obtained as a result of the screening had a salt tolerance enhancing function even at the plant level.

Example 7 Effect of Mangrove cDNA on Various Environmental Stresses (1) Heat Stress The cDNA shown in SEQ ID NO: 1 has been cloned.
SOLR introduced with pBluescript SK is 50 μg / ml
Kanamycin, 50 μg / ml ampicillin, 0.
The cells were cultured at 37 ° C. and 40 ° C. in a 2YT liquid medium containing 05 mM IPTG. The vector pBlu as a control
A similar study was conducted for SOLR into which escript SK was introduced. FIG. 6 shows the result. As is clear from FIG. 6, it was confirmed that the cDNA obtained as a result of the screening had a function of enhancing heat resistance.

(2) Osmotic stress The cDNA shown in SEQ ID NO: 1 has been cloned.
SOLR introduced with pBluescript SK is 50 μg / ml
Kanamycin, 50 μg / ml ampicillin, 0.
The cells were cultured in a 2YT liquid medium containing 05 mM IPTG until the logarithmic growth phase was reached, and a series of dilutions were prepared with the 2YT liquid medium, and 25 μl was added to 800 mM 2YT agar selection medium.
Each spot was spotted. After air drying until the liquid is dry, 37
Cultured overnight at ℃. The result is shown in FIG. As is clear from FIG. 7, the cD obtained as a result of the screening was used.
It was confirmed that NA has a function of enhancing osmotic pressure tolerance.

(3) Freezing stress The cDNA shown in SEQ ID NO: 1 has been cloned.
SOLR into which pBluescript SK was introduced was converted to 50 μg / ml kanamycin, 50 μg / ml ampicillin, 0.0
The cells were cultured in a 2YT liquid medium containing 5 mM IPTG until the logarithmic growth phase was reached.
Diluted to 11 ls / 25 μl. This was transferred to a plastic tube, and freezing with liquid nitrogen for 3 minutes and thawing at 37 ° C. for 10 minutes were repeated. A part (25 μl) of the cell suspension at the time of thawing was collected, and 50 μg of SOLR was collected.
/ Ml kanamycin, 50 μg / ml ampicillin, 0.05 mM IPTG on 2YT agar medium. For comparison, the vector pBluescript SK
A similar study was conducted for SOLR in which was introduced.
The result is shown in FIG. As is clear from FIG.
It was confirmed that the cDNA obtained as a result of the screening had a function of enhancing resistance to freezing stress.

Example 8 Molecular Evolution of Protein Having Activity to Improve Environmental Stress Tolerance A plasmid cloned as a region (amino acids 17-86), which is considered to be the minimum functional region of mangulin cDNA shown in FIG. 2, was used as a template. By performing PCR, the region considered to be the minimum functional region of mangulin (mangulin
rin core). As a primer, 5′-GCTCTGAGAACCGTC represented by SEQ ID NO: 65
TAGACTTAGATGAAGGTG-represented by 3 ′ and SEQ ID NO: 66
5 '-TCTCTCGTTCATCTCGAGCTATTACAGCTC-3' was used.
These primers add mangrincore to the start codon and stop codon, and the restriction enzymes (Xba1, Xho1)
It is designed so that it can be amplified by adding a site. In PCR, Ta is used as a DNA polymerase.
Kara Taq (Mg ²⁺ free buffer) (Takara Shuzo) was used.
The PCR reaction solution was prepared by adding MgCl ₂ to the attached buffer.
1.0 mM, MnCl ₂ 0.5 mM, dNPT Mix
was added to a concentration of 0.25 mM, and each primer was added thereto at 2 pmol / 10 μl, and template DNA was added at 10 pg / 10 μl. The temperature conditions for the reaction were 92 ° C. for 30 seconds, 50 ° C. for 30 seconds, 72 ° C.
By repeating this cycle 30 times at 90 ° C. for 90 seconds, DNA with random mutation introduced into mangrin core
A fragment was obtained. The resulting DNA fragment was cut with Xba1 and Xho1, and this was previously cut with Xba1 and Xho1.
(pBluescript SK). This is SOL
R, and selected using growth on an agar medium containing 450 mM NaCl as an index. Despite mutation in the base sequence, the salt-improving activity is maintained or enhanced. The core was selected. The nucleotide sequence of a part of the obtained clone was determined, and the activity of improving salt tolerance was evaluated by a spot test. As a result, as shown in FIG. 9, it was revealed that mutations occurred in the base sequences of the two clones (c-52, c-80). this house,
c-80 has a mutation in the amino acid sequence,
It was considered to be a factor that enhances the salt tolerance enhancing function of ngrincore. Moreover, as is clear from FIG. 10 showing a comparison of the salt stress tolerance enhancing function, it was found that it can be used for improving the function of the environmental stress tolerance gene.

[0066]

The present invention is an effective means for enhancing the resistance of various organisms to environmental stress. In particular, plants with enhanced resistance to environmental stress are promoted to grow in salt-damaged soils, cold regions, deserts, and oceans, where growth has been difficult so far, thereby increasing the production of agricultural products due to the expansion of agricultural land. In addition, plants with enhanced environmental stress tolerance not only lead to an increase in green space environments and desert greening, but also contribute to the suppression of global warming phenomena due to an increase in CO2 concentration on a global scale.

[0067]

[Sequence list]_{SEQUENCE LISTING  <110> JAPAN SCIENCE AND TECHNOLOGY CORPORATION <120> Screening of genes to give tolerance against environmental stress and the apprications <130> 12-130 <140> <141> <150> JP P1999-235910 <151> 1999-07-19 <150> JP P2000-85377 <151> 2000-03-24 <160> 66 <170> PatentIn Ver. 2.1 <210> 1 <211> 1018 <212> DNA <213> Bruguiera sexangula <220> <221> CDS <222> (42) .. (464) <400> 1 gtccaaacag ccagagagaa acgacaacat cgaccaagaa a atg gct ctt tca agc 56 Met Ala Leu Ser Ser 1 5 tct gct ctg aga acc gtc tct tct tct gtg aag gtg gtc ggc cct gca 104 Ser Ala Leu Arg Thr Val Ser Ser Ser Val Lys Val Val Gly Pro Ala 10 15 20 aga tca aag agt gct act gta ccc acc caa aca gta ttg cct ttc aag 152 Arg Ser Lys Ser Ala Thr Val Pro Thr Gln Thr Val Leu Pro Phe Lys 25 30 35 ttc aca aac ccg tcg tta ctc act cga tcg cta agc ttt tca tca aaa 200 Phe Thr Asn Pro Ser Leu Leu Thr Arg Ser Leu Ser Phe Ser Ser Lys 40 45 50 ggt tca agc ttt gac agc ttc tct gta ccc aaa aga tct ttt tct tgc 248 Gly Ser Ser Phe Asp Ser Phe Ser Val Pro Lys Arg Ser Phe Ser Cys 55 60 65 aga agc caa gcc act cca tct gat gat gcc tca aga ccc acc aaa gtt 296 Arg Ser Gln Ala Thr Pro Ser Asp Asp Ala Ser Arg Pro Thr Lys Val 70 75 80 85 caa gag ctg tgt gtg tat gag atg aac gag aga gat cgt gga agc cct 344 Gln Glu Leu Cys Val Tyr Glu Met Asn Glu Arg Asp Arg Gly Ser Pro 90 95 100 gct gtt ctc cgg ttg agc cag aaa cct gtt aat tct ctc ggc gat ctc 392 Ala Val Leu Arg Leu Ser Gln Lys Pro Val Asn Ser Leu Gly Asp Leu 105 110 115 gtg cct ttc agt aac aaa gtt tac agc gga gac ctg cag aag cga att 440 Val Pro Phe Ser Asn Lys Val Tyr Ser Gly Asp Leu Gln Lys Arg Ile 120 125 130 gga gta acc gca gaa tat gca tcc tgatccaaaa caagccagaa aaaaagggtg 494 Gly Val Thr Ala Glu Tyr Ala Ser 135 140 atcgctttga agcgatatat agcttttatt tcggtggcta tggtcacatt gctgtgcaag 554 gcgcatactt gacctacgag gacacgcacc ttgctgtgac gggcgggtcg ggcatatttg 614 aaggagtgtc tggtcaggtt aagctgcagc aactcgtgta ccctttcaag ctcttctaca 674 ctttctactt gcgaggcatc aaggacttgc cggaggagct tacgaagaag ccggttgagc 734 cccacccttc tgttgagccg atgccggcgg ccaaggcttg cgagccacat gccgttgttg 794 ctaatttcac cgattagtga ttaattgtcc ttttggggtt cggatgaact tgagttagct 854 tacagttgca caacgttatg gcgcgagaca cgagagggaa ccttagccat aagaaaatta 914 ataatctcac ggtgctttta ttttgattct tctattagtt gaatcgttaa tgaaagtgga 974 ccaaattggc tgttttacgt tttaaaaaaa aaaaaaaaaa aaaa 1018 <210> 2 <211> 141 <212> PRT <213> Bruguiera sexangula <400> 2 Met Ala Leu Ser Ser Ser Ala Leu Arg Thr Val Ser Ser Ser Val Lys 1 5 10 15 Val Val Gly Pro Ala Arg Ser Lys Ser Ala Thr Val Pro Thr Gln Thr 20 25 30 Val Leu Pro Phe Lys Phe Thr Asn Pro Ser Leu Leu Thr Arg Ser Leu 35 40 45 Ser Phe Ser Ser Lys Gly Ser Ser Phe Asp Ser Phe Ser Val Pro Lys 50 55 60 Arg Ser Phe Ser Cys Arg Ser Gln Ala Thr Pro Ser Asp Asp Ala Ser 65 70 75 80 Arg Pro Thr Lys Val Gln Glu Leu Cys Val Tyr Glu Met Asn Glu Arg 85 90 95 Asp Arg Gly Ser Pro Ala Val Leu Arg Leu Ser Gln Lys Pro Val Asn 100 105 110 Ser Leu Gly Asp Leu Val Pro Phe Ser Asn Lys Val Tyr Ser Gly Asp 115 120 125 Leu Gln Lys Arg Ile Gly Val Thr Ala Glu Tyr Ala Ser 130 135 140 <210> 3 <211> 2060 <212> DNA <213> Bruguiera sexangula <220> <221> CDS <222> (81) .. (1718) <400> 3 cgaaattcct ctactaacaa taccagatcc agtctagcgt ttcgattttc tgcttcacat 60 ttctgtttct ttgaccagaa atg gca atc gcg gct caa act ccg gac att ctc 113 Met Ala Ile Ala Ala Gln Thr Pro Asp Ile Leu 1 5 10 ggc gaa cgt cag tcc ggc cag gac gtc cgc act caa aat gtg gtg gca 161 Gly Glu Arg Gln Ser Gly Gln Asp Val Arg Thr Gln Asn Val Val Ala 15 20 25 tgt caa gcg gtt gcc aat att gtc aaa tct tca ctt ggt cct gtc gga 209 Cys Gln Ala Val Ala Asn Ile Val Lys Ser Ser Leu Gly Pro Val Gly 30 35 40 ctc gac aag atg cta gtg gat gat att ggt gat gta aca att aca aat 257 Leu Asp Lys Met Leu Val Asp Asp Ile Gly Asp Val Thr Ile Thr Asn 45 50 55 gat ggt gct acg att ctt aag atg tta gaa gta gag cat cct gca gca 305 Asp Gly Ala Thr Ile Leu Lys Met Leu Glu Val Glu His Pro Ala Ala 60 65 70 75 aag gtg ctc gtg gag ttg gct gag ctt caa gac cga gaa gtt gga gat 353 Lys Val Leu Val Glu Leu Ala Glu Leu Gln Asp Arg Glu Val Gly Asp 80 85 90 gga acc act tcg gtt gtc atc ata gca gct gag ttg ctc aag aga gca 401 Gly Thr Thr Ser Val Val Ile Ile Ala Ala Glu Leu Leu Lys Arg Ala 95 100 105 aat gat ctc gtg agg aat aag atc cac cca aca tca ata atc agt gga 449 Asn Asp Leu Val Arg Asn Lys Ile His Pro Thr Ser Ile Ile Ser Gly 110 115 120 tac agg ctt gct atg agg gaa gca tgc aag tat gtt gaa gag aaa ttg 497 Tyr Arg Leu Ala Met Arg Glu Ala Cys Lys Tyr Val Glu Glu Lys Leu 125 130 135 tca atg aag gtt gaa aag ctt gga aaa gat tct cta gta aac tgt gca 545 Ser Met Lys Val Glu Lys Leu Gly Lys Asp Ser Leu Val Asn Cys Ala 140 145 150 155 aag aca agc atg tcc tca aag ttg ata gct ggt gac agc gac ttc ttt 593 Lys Thr Ser Met Ser Ser Lys Leu Ile Ala Gly Asp Ser Asp Phe Phe 160 165 170 gca aat ttg gtt gta gat gct gta caa gca gta aag atg acc aat gca 641 Ala Asn Leu Val Val Asp Ala Val Gln Ala Val Lys Met Thr Asn Ala 175 180 185 cgg ggg gaa atc aaa tat cct atc aag agt ata aat att ttg aaa gct 689 Arg Gly Glu Ile Lys Tyr Pro Ile Lys Ser Ile Asn Ile Leu Lys Ala 190 195 200 cat gga aaa agt gca aga gat agc tgc ctt ttg aat ggc tat gct ctc 737 His Gly Lys Ser Ala Arg Asp Ser Cys Leu Leu Asn Gly Tyr Ala Leu 205 210 215 aat act ggt cgt gct gct caa ggg atg cct atg aga gtt gca cct gca 785 Asn Thr Gly Arg Ala Ala Gln Gly Met Pro Met Arg Val Ala Pro Ala 220 225 230 235 agg att gct tgt ctt gac ttt aat ctt cag aaa acg aag atg caa ttg 833 Arg Ile Ala Cys Leu Asp Phe Asn Leu Gln Lys Thr Lys Met Gln Leu 240 245 250 ggt gta caa gtc tta gtc act gat ccc agg gag ctt gaa aga att cgt 881 Gly Val Gln Val Leu Val Thr Asp Pro Arg Glu Leu Glu Arg Ile Arg 255 260 265 caa aga gaa gct gat atg aca aag gaa cgg att gag aaa ctc ctg aaa 929 Gln Arg Glu Ala Asp Met Thr Lys Glu Arg Ile Glu Lys Leu Leu Lys 270 275 280 gct gga gca aat gtt gtt cta acc aca aag gga att gat gac atg gca 977 Ala Gly Ala Asn Val Val Leu Thr Thr Lys Gly Ile Asp Asp Met Ala 285 290 295 ctt aaa tat ttt gtg gag gct ggg gct att gct gtg aga cgt gtt cgg 1025 Leu Lys Tyr Phe Val Glu Ala Gly Ala Ile Ala Val Arg Arg Val Arg 300 305 310 315 aaa gag gat atg cgc cat gtt gcc aag gca act ggt gca aca ctg gtt 1073 Lys Glu Asp Met Arg His Val Ala Lys Ala Thr Gly Ala Thr Leu Val 320 325 330 tca aca ttt gct gac atg gaa gga gag gaa aca ttt gat tca tca ctg 1121 Ser Thr Phe Ala Asp Met Glu Gly Glu Glu Thr Phe Asp Ser Ser Leu 335 340 345 ctt gga caa gct gaa gaa gtt gtg gag gag cgc att gct gat gac gat 1169 Leu Gly Gln Ala Glu Glu Val Val Glu Glu Arg Ile Ala Asp Asp Asp 350 355 360 gtg att atg ata aaa ggg aca aag act aca agt gcg gtt tcc ttg att 1217 Val Ile Met Ile Lys Gly Thr Lys Thr Thr Ser Ala Val Ser Leu Ile 365 370 375 ctt cgt ggt gca aat gac tat atg ctc gat gag atg gag cga gcc ctg 1265 Leu Arg Gly Ala Asn Asp Tyr Met Leu Asp Glu Met Glu Arg Ala Leu 380 385 390 395 cat gat gct tta tgt att gtc aag aga acc ctt gaa tct aat aca gta 1313 His Asp Ala Leu Cys Ile Val Lys Arg Thr Leu Glu Ser Asn Thr Val 400 405 410 gtt gca ggt gga ggt gct gtt gag gct gcc ttg tct gtg cac ttg gag 1361 Val Ala Gly Gly Gly Ala Val Glu Ala Ala Leu Ser Val His Leu Glu 415 420 425 tac ctc gct aca act ctt ggg tca cga gag cag tta gca ata gca gag 1409 Tyr Leu Ala Thr Thr Leu Gly Ser Arg Glu Gln Leu Ala Ile Ala Glu 430 435 440 ttt gca gaa tcc ttg ttg att ata cca aag gtt ctt gct gtc aat gct 1457 Phe Ala Glu Ser Leu Leu Ile Ile Pro Lys Val Leu Ala Val Asn Ala 445 450 455 gcc aaa gat gcc act gaa tta gct gca aaa ctc cgg gct tac cac cat 1505 Ala Lys Asp Ala Thr Glu Leu Ala Ala Lys Leu Arg Ala Tyr His His 460 465 470 475 aca gca caa aca aag gct gat aag aaa cat tta tca agc atg gga cta 1553 Thr Ala Gln Thr Lys Ala Asp Lys Lys His Leu Ser Ser Met Gly Leu 480 485 490 gac ctt tca aag ggg acc atc cga aac aac tta gaa gct gga gtc att 1601 Asp Leu Ser Lys Gly Thr Ile Arg Asn Asn Leu Glu Ala Gly Val Ile 495 500 505 gaa cct gca atg agc aaa ata aag ata att cag ttt gct act gaa gca 1649 Glu Pro Ala Met Ser Lys Ile Lys Ile Ile Gln Phe Ala Thr Glu Ala 510 515 520 gcc ata aca att ctt cga att gat gac atg atc aag ctt gtc aag gat 1697 Ala Ile Thr Ile Leu Arg Ile Asp Asp Met Ile Lys Leu Val Lys Asp 525 530 535 gag act cag aat gaa gag gaa tagatgcaga ctcttgtaag ctgcctccct 1748 Glu Thr Gln Asn Glu Glu Glu 540 545 tttgttttca aatttgtgtc ccttgcgagc tggaggaaag ggggggtgtt tatgtggtgt 1808 tttcagtggt tttaattttt caaggagctc gcggcctgtg tactttaggt tagagtccat 1868 ccaaggggtg tttattggat aatgcctaag ctgtttctcg tctattagta ggctggtagt 1928 tccactgagt tctcatccca attaaaagaa tgagatcaaa gggtcctaaa ttcgtactca 1988 ttggtgcacg atttgtttct gacaagcata agacttgacc ctctctatca caataaaaaa 2048 aaaaaaaaaa aa 2060 <210> 4 <211> 546 <212> PRT <213> Bruguiera sexangula <400> 4 Met Ala Ile Ala Ala Gln Thr Pro Asp Ile Leu Gly Glu Arg Gln Ser 1 5 10 15 Gly Gln Asp Val Arg Thr Gln Asn Val Val Ala Cys Gln Ala Val Ala 20 25 30 Asn Ile Val Lys Ser Ser Leu Gly Pro Val Gly Leu Asp Lys Met Leu 35 40 45 Val Asp Asp Ile Gly Asp Val Thr Ile Thr Asn Asp Gly Ala Thr Ile 50 55 60 Leu Lys Met Leu Glu Val Glu His Pro Ala Ala Lys Val Leu Val Glu 65 70 75 80 Leu Ala Glu Leu Gln Asp Arg Glu Val Gly Asp Gly Thr Thr Ser Val 85 90 95 Val Ile Ile Ala Ala Glu Leu Leu Lys Arg Ala Asn Asp Leu Val Arg 100 105 110 Asn Lys Ile His Pro Thr Ser Ile Ile Ser Gly Tyr Arg Leu Ala Met 115 120 125 Arg Glu Ala Cys Lys Tyr Val Glu Glu Lys Leu Ser Met Lys Val Glu 130 135 140 Lys Leu Gly Lys Asp Ser Leu Val Asn Cys Ala Lys Thr Ser Met Ser 145 150 155 160 Ser Lys Leu Ile Ala Gly Asp Ser Asp Phe Phe Ala Asn Leu Val Val 165 170 175 Asp Ala Val Gln Ala Val Lys Met Thr Asn Ala Arg Gly Glu Ile Lys 180 185 190 Tyr Pro Ile Lys Ser Ile Asn Ile Leu Lys Ala His Gly Lys Ser Ala 195 200 205 Arg Asp Ser Cys Leu Leu Asn Gly Tyr Ala Leu Asn Thr Gly Arg Ala 210 215 220 Ala Gln Gly Met Pro Met Arg Val Ala Pro Ala Arg Ile Ala Cys Leu 225 230 235 240 Asp Phe Asn Leu Gln Lys Thr Lys Met Gln Leu Gly Val Gln Val Leu 245 250 255 Val Thr Asp Pro Arg Glu Leu Glu Arg Ile Arg Gln Arg Glu Ala Asp 260 265 270 Met Thr Lys Glu Arg Ile Glu Lys Leu Leu Lys Ala Gly Ala Asn Val 275 280 285 Val Leu Thr Thr Lys Gly Ile Asp Asp Met Ala Leu Lys Tyr Phe Val 290 295 300 Glu Ala Gly Ala Ile Ala Val Arg Arg Val Arg Lys Glu Asp Met Arg 305 310 315 320 His Val Ala Lys Ala Thr Gly Ala Thr Leu Val Ser Thr Phe Ala Asp 325 330 335 Met Glu Gly Glu Glu Thr Phe Asp Ser Ser Leu Leu Gly Gln Ala Glu 340 345 350 Glu Val Val Glu Glu Arg Ile Ala Asp Asp Asp Val Ile Met Ile Lys 355 360 365 Gly Thr Lys Thr Thr Ser Ala Val Ser Leu Ile Leu Arg Gly Ala Asn 370 375 380 Asp Tyr Met Leu Asp Glu Met Glu Arg Ala Leu His Asp Ala Leu Cys 385 390 395 400 Ile Val Lys Arg Thr Leu Glu Ser Asn Thr Val Val Ala Gly Gly Gly 405 410 415 Ala Val Glu Ala Ala Leu Ser Val His Leu Glu Tyr Leu Ala Thr Thr 420 425 430 Leu Gly Ser Arg Glu Gln Leu Ala Ile Ala Glu Phe Ala Glu Ser Leu 435 440 445 Leu Ile Ile Pro Lys Val Leu Ala Val Asn Ala Ala Lys Asp Ala Thr 450 455 460 Glu Leu Ala Ala Lys Leu Arg Ala Tyr His His Thr Ala Gln Thr Lys 465 470 475 480 Ala Asp Lys Lys His Leu Ser Ser Met Gly Leu Asp Leu Ser Lys Gly 485 490 495 Thr Ile Arg Asn Asn Leu Glu Ala Gly Val Ile Glu Pro Ala Met Ser 500 505 510 Lys Ile Lys Ile Ile Gln Phe Ala Thr Glu Ala Ala Ile Thr Ile Leu 515 520 525 Arg Ile Asp Asp Met Ile Lys Leu Val Lys Asp Glu Thr Gln Asn Glu 530 535 540 Glu Glu 545 <210> 5 <211> 588 <212> DNA <213> Bruguiera sexangula <220> <221> CDS <222> (26) .. (262) <400> 5 gaaaaacaaa gcaatctcct gaagg atg tct tgc tgt ggt gga aac tgt ggc 52 Met Ser Cys Cys Gly Gly Asn Cys Gly 1 5 tgc gga gca agc tgc aat tgc ggc aac ggc tgt gga ggg tgc aag atg 100 Cys Gly Ala Ser Cys Asn Cys Gly Asn Gly Cys Gly Gly Cys Lys Met 10 15 20 25 tac cca gac atg ggc ttc gcc gag aag acc act acc gag act ctg gtt 148 Tyr Pro Asp Met Gly Phe Ala Glu Lys Thr Thr Thr Glu Thr Leu Val 30 35 40 ctc ggc gtg ggg cct gag agg gcc cac ttt gag gga gcc gag atg ggc 196 Leu Gly Val Gly Pro Glu Arg Ala His Phe Glu Gly Ala Glu Met Gly 45 50 55 gtg ccg gcc gag aac gga ggc tgc aag tgc gga agt aac tgc acc tgc 244 Val Pro Ala Glu Asn Gly Gly Cys Lys Cys Gly Ser Asn Cys Thr Cys 60 65 70 gac ccc tgc act tgt aaa tgaggggaaa gtgacaggga aggtccgatc 292 Asp Pro Cys Thr Cys Lys 75 tattattagt ctatatgtgt gtgttgggag tcttgcttac aataaaccag tcatgccttg 352 cgtttcctcc atgcgcagat cttaggtttt aggatatctc tgtggtttct ccaagctatg 412 gattttcagt gtctagtttt cctgtattac aaggatagtt tataaccgta tatgcatggt 472 cggaatcctt ccaaccattt cgtttgtcta aatatatata tgtgtgtgtg tgtgtgtgtt 532 tgatgggaaa gtgagcttct ttatgtttta tgactaaaaa aaaaaaaaaa aaaaaa 588 <210> 6 <211> 79 <212> PRT <213> Bruguiera sexangula <400> 6 Met Ser Cys Cys Gly Gly Asn Cys Gly Cys Gly Ala Ser Cys Asn Cys 1 5 10 15 Gly Asn Gly Cys Gly Gly Cys Lys Met Tyr Pro Asp Met Gly Phe Ala 20 25 30 Glu Lys Thr Thr Thr Glu Thr Leu Val Leu Gly Val Gly Pro Glu Arg 35 40 45 Ala His Phe Glu Gly Ala Glu Met Gly Val Pro Ala Glu Asn Gly Gly 50 55 60 Cys Lys Cys Gly Ser Asn Cys Thr Cys Asp Pro Cys Thr Cys Lys 65 70 75 <210> 7 <211> 1280 <212> DNA <213> Bruguiera sexangula <220> <221> CDS <222> (1) .. (1002) <400> 7 att gaa ggg gaa gtg gtg gaa gtc caa att gat cgg ccg gcg gtg acc 48 Ile Glu Gly Glu Val Val Glu Val Gln Ile Asp Arg Pro Ala Val Thr 1 5 10 15 ggc gcc gcg tcc aag acg ggg aaa ttg acg cta aag acg acg gag atg 96 Gly Ala Ala Ser Lys Thr Gly Lys Leu Thr Leu Lys Thr Thr Glu Met 20 25 30 gag acg gtg tac gat ttg ggg gcg aaa atg ata gag gca ttg ggg aag 144 Glu Thr Val Tyr Asp Leu Gly Ala Lys Met Ile Glu Ala Leu Gly Lys 35 40 45 gaa aag gtg cag agt ggg gat gtt att gca att gac aag gcg tcc ggc 192 Glu Lys Val Gln Ser Gly Asp Val Ile Ala Ile Asp Lys Ala Ser Gly 50 55 60 aaa att aca aag ctt ggg cgt tca ttt tcg cgg tct agg gat tac gat 240 Lys Ile Thr Lys Leu Gly Arg Ser Phe Ser Arg Ser Arg Asp Tyr Asp 65 70 75 80 gcc atg gga cca cag gtg aag ttt gtt cag tgc cct gat ggg gag ctg 288 Ala Met Gly Pro Gln Val Lys Phe Val Gln Cys Pro Asp Gly Glu Leu 85 90 95 cag aag agg aaa gag gtc gtg cat tgt gtc tca ctg cac gag att gat 336 Gln Lys Arg Lys Glu Val Val His Cys Val Ser Leu His Glu Ile Asp 100 105 110 gtt atc aat agc aga aca cag ggg ttt ctt gct ctt ttc acc ggg gat 384 Val Ile Asn Ser Arg Thr Gln Gly Phe Leu Ala Leu Phe Thr Gly Asp 115 120 125 act ggt gaa atc cgt gcg gag gtg agg gaa caa att gac aca aag gtg 432 Thr Gly Glu Ile Arg Ala Glu Val Arg Glu Gln Ile Asp Thr Lys Val 130 135 140 gct gaa tgg aga gag gaa ggg aaa gca gag att gtg cca ggt gtc ctc 480 Ala Glu Trp Arg Glu Glu Gly Lys Ala Glu Ile Val Pro Gly Val Leu 145 150 155 160 ttt att gat gag gtc cac atg ctt gac att gag tgc ttc tca ttt ctg 528 Phe Ile Asp Glu Val His Met Leu Asp Ile Glu Cys Phe Ser Phe Leu 165 170 175 aat cgt gct ctt gag aat gag atg gcg cca ata tta gtt gtt gct acc 576 Asn Arg Ala Leu Glu Asn Glu Met Ala Pro Ile Leu Val Val Ala Thr 180 185 190 aac aga ggg atc acc aca atc aga ggc aca aat tac aaa tct cct cat 624 Asn Arg Gly Ile Thr Thr Ile Arg Gly Thr Asn Tyr Lys Ser Pro His 195 200 205 ggg att cca ata gat ctc ctt gat cga cta ctc att atc aca act caa 672 Gly Ile Pro Ile Asp Leu Leu Asp Arg Leu Leu Ile Ile Thr Thr Gln 210 215 220 cct tac aca aag gat gaa att cgt aag att ctg gat atc aga tgt cag 720 Pro Tyr Thr Lys Asp Glu Ile Arg Lys Ile Leu Asp Ile Arg Cys Gln 225 230 235 240 gaa gaa gat gtg gag atg gct gaa gag gca aag gct ttg tta aca cat 768 Glu Glu Asp Val Glu Met Ala Glu Glu Ala Lys Ala Leu Leu Thr His 245 250 255 att ggg gca gaa aca tcc ttg aga tat gcc atc cat ctc att act gct 816 Ile Gly Ala Glu Thr Ser Leu Arg Tyr Ala Ile His Leu Ile Thr Ala 260 265 270 gca gca ttg gca tgc cag aag cga aag gga aag ctt gtg gaa act gag 864 Ala Ala Leu Ala Cys Gln Lys Arg Lys Gly Lys Leu Val Glu Thr Glu 275 280 285 gac att agt cga gct tac aat ctg ttt ctt gat gta aag aga tct aca 912 Asp Ile Ser Arg Ala Tyr Asn Leu Phe Leu Asp Val Lys Arg Ser Thr 290 295 300 cag tac cta ata gag tat cag aat cag tac atg ttt aat gag gca ccg 960 Gln Tyr Leu Ile Glu Tyr Gln Asn Gln Tyr Met Phe Asn Glu Ala Pro 305 310 315 320 gta gga gaa ggg gac gaa gaa ggg gcc aat gcc atg ctt tct 1002 Val Gly Glu Gly Asp Glu Glu Gly Ala Asn Ala Met Leu Ser 325 330 tgaagggcca taagctatgg agtctttgtg aaacccttct ccctacttta ttcgcagcac 1062 gagccctgaa atgaagaaca atggtagact tggatcccac cttggccctt atgtatgtct 1122 tctggaattg aaaaaagagt ccaagaaatt tgaatttcat gaaattggag aactgaactg 1182 tgcttactaa attgctactt tgcaagtaat gatagggcac tcacgcttga ctggctaagt 1242 atttatgttt ttatcatcaa aaaaaaaaaa aaaaaaaa 1280 <210> 8 <211> 334 <212> PRT <213> Bruguiera sexangula <400> 8 Ile Glu Gly Glu Val Val Glu Val Gln Ile Asp Arg Pro Ala Val Thr 1 5 10 15 Gly Ala Ala Ser Lys Thr Gly Lys Leu Thr Leu Lys Thr Thr Glu Met 20 25 30 Glu Thr Val Tyr Asp Leu Gly Ala Lys Met Ile Glu Ala Leu Gly Lys 35 40 45 Glu Lys Val Gln Ser Gly Asp Val Ile Ala Ile Asp Lys Ala Ser Gly 50 55 60 Lys Ile Thr Lys Leu Gly Arg Ser Phe Ser Arg Ser Arg Asp Tyr Asp 65 70 75 80 Ala Met Gly Pro Gln Val Lys Phe Val Gln Cys Pro Asp Gly Glu Leu 85 90 95 Gln Lys Arg Lys Glu Val Val His Cys Val Ser Leu His Glu Ile Asp 100 105 110 Val Ile Asn Ser Arg Thr Gln Gly Phe Leu Ala Leu Phe Thr Gly Asp 115 120 125 Thr Gly Glu Ile Arg Ala Glu Val Arg Glu Gln Ile Asp Thr Lys Val 130 135 140 Ala Glu Trp Arg Glu Glu Gly Lys Ala Glu Ile Val Pro Gly Val Leu 145 150 155 160 Phe Ile Asp Glu Val His Met Leu Asp Ile Glu Cys Phe Ser Phe Leu 165 170 175 Asn Arg Ala Leu Glu Asn Glu Met Ala Pro Ile Leu Val Val Ala Thr 180 185 190 Asn Arg Gly Ile Thr Thr Ile Arg Gly Thr Asn Tyr Lys Ser Pro His 195 200 205 Gly Ile Pro Ile Asp Leu Leu Asp Arg Leu Leu Ile Ile Thr Thr Gln 210 215 220 Pro Tyr Thr Lys Asp Glu Ile Arg Lys Ile Leu Asp Ile Arg Cys Gln 225 230 235 240 Glu Glu Asp Val Glu Met Ala Glu Glu Ala Lys Ala Leu Leu Thr His 245 250 255 Ile Gly Ala Glu Thr Ser Leu Arg Tyr Ala Ile His Leu Ile Thr Ala 260 265 270 Ala Ala Leu Ala Cys Gln Lys Arg Lys Gly Lys Leu Val Glu Thr Glu 275 280 285 Asp Ile Ser Arg Ala Tyr Asn Leu Phe Leu Asp Val Lys Arg Ser Thr 290 295 300 Gln Tyr Leu Ile Glu Tyr Gln Asn Gln Tyr Met Phe Asn Glu Ala Pro 305 310 315 320 Val Gly Glu Gly Asp Glu Glu Gly Ala Asn Ala Met Leu Ser 325 330 <210> 9 <211> 420 <212> DNA <213> Bruguiera sexangula <220> <221> CDS <222> (27) .. (194) <400> 9 cgaaagtata aagtgatcgg cgagcg atg ggt cac tct aac gtc tgg aac tct 53 Met Gly His Ser Asn Val Trp Asn Ser 1 5 cac ccc aag aac tac ggc cct ggt tcc cgc gcc tgt cgg gtg tgt ggg 101 His Pro Lys Asn Tyr Gly Pro Gly Ser Arg Ala Cys Arg Val Cys Gly 10 15 20 25 aat ccg cac ggg ttg atc agg aag tac gga ctc atg tgc tgc aga cag 149 Asn Pro His Gly Leu Ile Arg Lys Tyr Gly Leu Met Cys Cys Arg Gln 30 35 40 tgc ttc cgt agc aat gcc aag gaa att ggc ttc att aag tac cgc 194 Cys Phe Arg Ser Asn Ala Lys Glu Ile Gly Phe Ile Lys Tyr Arg 45 50 55 tgaatgatat cgatatggcc cagaatggcc tgtggcggtg cgtgttcgat ttcagtagtt 254 cccctctttc ggatgagctt taggacaatg ttctctttag tttatgtatt gttgaacttg 314 gactgatgtt gaactaacga tattctggaa tcatttgata tttcgagagt ttattatttt 374 gatcatcatc ctcttgcttc tctgcttaaa aaaaaaaaaa aaaaaa 420 <210> 10 <211> 56 <212> PRT <213> Bruguiera sexangula <400> 10 Met Gly His Ser Asn Val Trp Asn Ser His Pro Lys Asn Tyr Gly Pro 1 5 10 15 Gly Ser Arg Ala Cys Arg Val Cys Gly Asn Pro His Gly Leu Ile Arg 20 25 30 Lys Tyr Gly Leu Met Cys Cys Arg Gln Cys Phe Arg Ser Asn Ala Lys 35 40 45 Glu Ile Gly Phe Ile Lys Tyr Arg 50 55 <210> 11 <211> 1664 <212> DNA <213> Bruguiera sexangula <220> <221> CDS <222> (34). . (1380) <400> 11 tctctctttta caggttaaaag ctaagacttt data
atg ggt aag gag aag att cac 54
Met Gly Lys Glu Lys Ile His
15 att aac att gtg gtt att ggc cat gtc
gac tcc gga aag tca acc aca 102 Ile Asn Ile Val Val Ile Gly His Val
Asp Ser Gly Lys Ser Thr Thr 10 15
20 act ggc cac tttg att tac aag ctt gga
ggt atc gac aag cgt gtg att 150 Thr Gly His Leu Ile Tyr Lys Leu Gly
Gly Ile Asp Lys Arg Val Ile 25 30
35 gag agg ttt gag aag gaa gct gct gag
atg aac aag agg tca tttc aag 198 Glu Arg Phe Glu Lys Glu Ala Ala Glu
Met Asn Lys Arg Ser Phe Lys 40 45
50 55 tat gcc tgg gtg ctt gac aag ctg aag
gct gag cgt gag cgt ggt atc 246 Tyr Ala Trp Val Leu Asp Lys Leu Lys
Ala Glu Arg Glu Arg Gly Ile 60
65 70 acc att gat att gcc ttg tgg aag ttc gag aca acc aaa tat tac tgc 294 Thr Ile Asp Ile Ala Leu Trp Lys Phe Glu Thr Thr Lys Tyr Tyr Cys 75 80 85 acg gtc att gat gct cct gga cat cgt gac ttt att aag aat atg atc 342 Thr Val Ile Asp Ala Pro Gly His Arg Asp Phe Ile Lys Asn Met Ile 90 95 100 acc ggg act tcc caa gct gac tgt gct gtc ctc atc att gac tct acc 390 Thr Gly Thr Ser Gln Ala Asp Cys Ala Val Leu Ile Ile Asp Ser Thr 105 110 115 act ggt ggc ttt gag gct ggt atc tct aaa gat ggt cag acc cgc gag 438 Thr Gly Gly Phe Glu Ala Gly Ile Ser Lys Asp Gly Gln Thr Arg Glu 120 125 130 135 cat gcc ctg ctt gcc ttc acc ctt ggt gtt aag caa atg att tgc tgc 486 His Ala Leu Leu Ala Phe Thr Leu Gly Val Lys Gln Met Ile Cys Cys 140 145 150 tgc aac aag atg gat gct acc act tcc aag tat tct aag gca aga tat 534 Cys Asn Lys Met Asp Ala Thr Thr Ser Lys Tyr Ser Lys Ala Arg Tyr 155 160 165 gat gaa att gtt aag gaa gtg tca tcc tac ttg aag aag gtt ggt tac 582 Asp Glu Ile Val Lys Glu Val Ser Ser Tyr Leu Lys Lys Val Gly Tyr 170 175 180 aac cca gag aag att cct ttt gtc ccc ata tct gga ttt gag ggt gac 630 Asn Pro Glu Lys Ile Pro Phe Val Pro Ile Ser Gly Phe Glu Gly Asp 185 190 195 aac atg att gag aga tcc acc aac ctt gac tgg tac aag ggc cca act 678 Asn Met Ile Glu Arg Ser Thr Asn Leu Asp Trp Tyr Lys Gly Pro Thr 200 205 210 215 ctt ctt gag gcc ctg gac atg atc cag gag cca aag agg cca tca gat 726 Leu Leu Glu Ala Leu Asp Met Ile Gln Glu Pro Lys Arg Pro Ser Asp 220 225 230 aag ccc ctc cgt ctc cca ctt cag gat gtg tac aag att ggt ggt att 774 Lys Pro Leu Arg Leu Pro Leu Gln Asp Val Tyr Lys Ile Gly Gly Ile 235 240 245 ggg aca gtc cca gtg ggt cgt gtt gaa act ggt gtc ctg aag cct gga 822 Gly Thr Val Pro Val Gly Arg Val Glu Thr Gly Val Leu Lys Pro Gly 250 255 260 atg gtt gtt act ttt ggt ccc tca gga ctg acc act gaa gtt aag tct 870 Met Val Val Thr Phe Gly Pro Ser Gly Leu Thr Thr Glu Val Lys Ser 265 270 275 gtg gag atg cac cat gaa gct ctc caa gag gct ctt ccc gga gac aac 918 Val Glu Met His His Glu Ala Leu Gln Glu Ala Leu Pro Gly Asp Asn 280 285 290 295 gtt ggc ttc aat gtt aag aat gtt tcc gtg aag gat ctt aag cgg ggt 966 Val Gly Phe Asn Val Lys Asn Val Ser Val Lys Asp Leu Lys Arg Gly 300 305 310 tat gtt gcc tca aac tcc aag gat gat cct gcc aag gag gca tct agc 1014 Tyr Val Ala Ser Asn Ser Lys Asp Asp Pro Ala Lys Glu Ala Ser Ser 315 320 325 ttc acc tcc caa gtt atc atc atg aac cac cct ggt cag att gga aat 1062 Phe Thr Ser Gln Val Ile Ile Met Asn His Pro Gly Gln Ile Gly Asn 330 335 340 ggt tat gcc cct gtt ctg gat tgc cac acc tct cac att gct gtc aag 1110 Gly Tyr Ala Pro Val Leu Asp Cys His Thr Ser His Ile Ala Val Lys 345 350 355 ttt tct gag atc ctc aca aag att gat agg cga tct ggc aag gag ctt 1158 Phe Ser Glu Ile Leu Thr Lys Ile Asp Arg Arg Ser Gly Lys Glu Leu 360 365 370 375 gaa aag gag ccc aag ttc ttg aag aat ggt gat gct ggg ttc gtg aag 1206 Glu Lys Glu Pro Lys Phe Leu Lys Asn Gly Asp Ala Gly Phe Val Lys 380 385 390 atg att ccg acc aag cct atg gtg gtg gaa act ttc tcc gag tat cct 1254 Met Ile Pro Thr Lys Pro Met Val Val Glu Thr Phe Ser Glu Tyr Pro 395 400 405 ccg ctt ggt aga ttt gcc gtc agg gac atg cgc cag act gtt gca gtg 1302 Pro Leu Gly Arg Phe Ala Val Arg Asp Met Arg Gln Thr Val Ala Val 410 415 420 gga gtc atc aag agt gtc gag aaa aag gaa cct tct gga gct aag gtg 1350 Gly Val Ile Lys Ser Val Glu Lys Lys Glu Pro Ser Gly Ala Lys Val 425 430 435 act aaa tct gct gcc aag aag ggt ggc aaa tgaaccgtgc aagtcagagt 1400 Thr Lys Ser Ala Ala Lys Lys Gly Gly Lys 440 445 tgatgtagat gaaggctatt ggaagaataa agactgggcc ctggttagcg gtctaattat 1460 tggatgttca gcagttggtt tcgagaacta cagtttcaat tcagcgccat catcacggag 1520 ctgttgttcc cagaattggg ttcttgaccg tcggtggcat tggctgttgg tttgagtgac 1580 ttctttgtgt catgtttaga ctttatcgga tttgctattt cataaagcgg cttgggaatt 1640 ttaaaaaaaa aaaaaaaaaa aaaa 1664 <210> 12 <211> 449 <212> PRT <213> Bruguiera sexangula <400> 12 Met Gly Lys Glu Lys Ile His Ile Asn Ile Val Val Ile Gly His Val 1 5 10 15 Asp Ser Gly Lys Ser Thr Thr Thr Gly His Leu Ile Tyr Lys Leu Gly 20 25 30 Gly Ile Asp Lys Arg Val Ile Glu Arg Phe Glu Lys Glu Ala Ala Glu 35 40 45 Met Asn Lys Arg Ser Phe Lys Tyr Ala Trp Val Leu Asp Lys Leu Lys 50 55 60 Ala Glu Arg Glu Arg Gly Ile Thr Ile Asp Ile Ala Leu Trp Lys Phe 65 70 75 80 Glu Thr Thr Lys Tyr Tyr Cys Thr Val Ile Asp Ala Pro Gly His Arg 85 90 95 Asp Phe Ile Lys Asn Met Ile Thr Gly Thr Ser Gln Ala Asp Cys Ala 100 105 110 Val Leu Ile Ile Asp Ser Thr Thr Gly Gly Phe Glu Ala Gly Ile Ser 115 120 125 Lys Asp Gly Gln Thr Arg Glu His Ala Leu Leu Ala Phe Thr Leu Gly 130 135 140 Val Lys Gln Met Ile Cys Cys Cys Asn Lys Met Asp Ala Thr Thr Ser 145 150 155 160 Lys Tyr Ser Lys Ala Arg Tyr Asp Glu Ile Val Lys Glu Val Ser Ser 165 170 175 Tyr Leu Lys Lys Val Gly Tyr Asn Pro Glu Lys Ile Pro Phe Val Pro 180 185 190 Ile Ser Gly Phe Glu Gly Asp Asn Met Ile Glu Arg Ser Thr Asn Leu 195 200 205 Asp Trp Tyr Lys Gly Pro Thr Leu Leu Glu Ala Leu Asp Met Ile Gln 210 215 220 Glu Pro Lys Arg Pro Ser Asp Lys Pro Leu Arg Leu Pro Leu Gln Asp 225 230 235 240 Val Tyr Lys Ile Gly Gly Ile Gly Thr Val Pro Val Gly Arg Val Glu 245 250 255 Thr Gly Val Leu Lys Pro Gly Met Val Val Thr Phe Gly Pro Ser Gly 260 265 270 Leu Thr Thr Glu Val Lys Ser Val Glu Met His His Glu Ala Leu Gln 275 280 285 Glu Ala Leu Pro Gly Asp Asn Val Gly Phe Asn Val Lys Asn Val Ser 290 295 300 Val Lys Asp Leu Lys Arg Gly Tyr Val Ala Ser Asn Ser Lys Asp Asp 305 310 315 320 Pro Ala Lys Glu Ala Ser Ser Phe Thr Ser Gln Val Ile Ile Met Asn 325 330 335 His Pro Gly Gln Ile Gly Asn Gly Tyr Ala Pro Val Leu Asp Cys His 340 345 350 Thr Ser His Ile Ala Val Lys Phe Ser Glu Ile Leu Thr Lys Ile Asp 355 360 365 Arg Arg Ser Gly Lys Glu Leu Glu Lys Glu Pro Lys Phe Leu Lys Asn 370 375 380 Gly Asp Ala Gly Phe Val Lys Met Ile Pro Thr Lys Pro Met Val Val 385 390 395 400 Glu Thr Phe Ser Glu Tyr Pro Pro Leu Gly Arg Phe Ala Val Arg Asp 405 410 415 Met Arg Gln Thr Val Ala Val Gly Val Ile Lys Ser Val Glu Lys Lys 420 425 430 Glu Pro Ser Gly Ala Lys Val Thr Lys Ser Ala Ala Lys Lys Gly Gly 435 440 445 Lys  <210> 13 <211> 770 <212> DNA <213> Bruguiera sexangula <220> <221> CDS <222> (2) .. (769) <400> 13 c gat gat atg gac gag gcc aca ccc acc ttt gtt tgg ggc acc aat atc 49 Asp Asp Met Asp Glu Ala Thr Pro Thr Phe Val Trp Gly Thr Asn Ile 1 5 10 15 agc gtg cag gat gtc aag gcc gct att cag atg ttt ttg aag cac ttc 97 Ser Val Gln Asp Val Lys Ala Ala Ile Gln Met Phe Leu Lys His Phe 20 25 30 agg gat agt aat cag agt caa agg aac gag att ttt gaa gaa ggg aag 145 Arg Asp Ser Asn Gln Ser Gln Arg Asn Glu Ile Phe Glu Glu Gly Lys 35 40 45 tac gtg aaa gcg ata cat aag gtt ctt gaa gtt gaa gga gag tcg ctt 193 Tyr Val Lys Ala Ile His Lys Val Leu Glu Val Glu Gly Glu Ser Leu 50 55 60 gat gtt gat gct cgt gat gtg ttt gat tat gat tct gat ttg tat gcc 241 Asp Val Asp Ala Arg Asp Val Phe Asp Tyr Asp Ser Asp Leu Tyr Ala 65 70 75 80 aag atg att cgg tac cca ctt gag gtt ttg gcc att ttc gac att gtt 289 Lys Met Ile Arg Tyr Pro Leu Glu Val Leu Ala Ile Phe Asp Ile Val 85 90 95 ttg atg gat att gtg agt ttg atc aac cct ttg ttt gag aaa cat gta 337 Leu Met Asp Ile Val Ser Leu Ile Asn Pro Leu Phe Glu Lys His Val 100 105 110 caa gtc agg att ttc aat ctt aag acc tcg att aca atg aga aat ctc 385 Gln Val Arg Ile Phe Asn Leu Lys Thr Ser Ile Thr Met Arg Asn Leu 115 120 125 aac cct tct gat atc gaa aag atg gtg tca ttg aag gga atg ata att 433 Asn Pro Ser Asp Ile Glu Lys Met Val Ser Leu Lys Gly Met Ile Ile 130 135 140 cgg tgt agt tcc ata ata ccg gag atc agg gaa gca gta ttt aga tgc 481 Arg Cys Ser Ser Ile Ile Pro Glu Ile Arg Glu Ala Val Phe Arg Cys 145 150 155 160 ctt gtt tgt ggc tac ttc tct gat ccc atc gtt gtg gat aga gga cgg 529 Leu Val Cys Gly Tyr Phe Ser Asp Pro Ile Val Val Asp Arg Gly Arg 165 170 175 ata agt gaa cct aaa gca tgc ttg aaa gag gaa tgt ctt act aag aac 577 Ile Ser Glu Pro Lys Ala Cys Leu Lys Glu Glu Cys Leu Thr Lys Asn 180 185 190 tcc atg aca cta gtt cac aat cgt tgc agg ttt gct gat aag cag att 625 Ser Met Thr Leu Val His Asn Arg Cys Arg Phe Ala Asp Lys Gln Ile 195 200 205 gtg agg ctc cag gag aca cct gac gag atc cct gaa gga gga aca cca 673 Val Arg Leu Gln Glu Thr Pro Asp Glu Ile Pro Glu Gly Gly Thr Pro 210 215 220 cac acg gtg agc tta ttg atg cat gac aag ctg gta gat gct gga aag 721 His Thr Val Ser Leu Leu Met His Asp Lys Leu Val Asp Ala Gly Lys 225 230 235 240 cca ggt gac agg gtt gag gtc act gga att tat agg gct atg agt gtt a 770 Pro Gly Asp Arg Val Glu Val Thr Gly Ile Tyr Arg Ala Met Ser Val 245 250 255 <210> 14 <211> 256 <212> PRT <213> Bruguiera sexangula <400> 14 Asp Asp Met Asp Glu Ala Thr Pro Thr Phe Val Trp Gly Thr Asn Ile 1 5 10 15 Ser Val Gln Asp Val Lys Ala Ala Ile Gln Met Phe Leu Lys His Phe 20 25 30 Arg Asp Ser Asn Gln Ser Gln Arg Asn Glu Ile Phe Glu Glu Gly Lys 35 40 45 Tyr Val Lys Ala Ile His Lys Val Leu Glu Val Glu Gly Glu Ser Leu 50 55 60 Asp Val Asp Ala Arg Asp Val Phe Asp Tyr Asp Ser Asp Leu Tyr Ala 65 70 75 80 Lys Met Ile Arg Tyr Pro Leu Glu Val Leu Ala Ile Phe Asp Ile Val 85 90 95 Leu Met Asp Ile Val Ser Leu Ile Asn Pro Leu Phe Glu Lys His Val 100 105 110 Gln Val Arg Ile Phe Asn Leu Lys Thr Ser Ile Thr Met Arg Asn Leu 115 120 125 Asn Pro Ser Asp Ile Glu Lys Met Val Ser Leu Lys Gly Met Ile Ile 130 135 140 Arg Cys Ser Ser Ile Ile Pro Glu Ile Arg Glu Ala Val Phe Arg Cys 145 150 155 160 Leu Val Cys Gly Tyr Phe Ser Asp Pro Ile Val Val Asp Arg Gly Arg 165 170 175 Ile Ser Glu Pro Lys Ala Cys Leu Lys Glu Glu Cys Leu Thr Lys Asn 180 185 190 Ser Met Thr Leu Val His Asn Arg Cys Arg Phe Ala Asp Lys Gln Ile 195 200 205 Val Arg Leu Gln Glu Thr Pro Asp Glu Ile Pro Glu Gly Gly Thr Pro 210 215 220 His Thr Val Ser Leu Leu Met His Asp Lys Leu Val Asp Ala Gly Lys 225 230 235 240 Pro Gly Asp Arg Val Glu Val Thr Gly Ile Tyr Arg Ala Met Ser Val 245 250 255 <210> 15 <211> 846 <212> DNA <213> Mesembryanthemum crystallinum <220> <221> CDS <222> (39) .. (530) <400> 15 caaattttct ttgctgaatc gaatctacaa aatacctg atg ggt cag gtt ctt gac 56 Met Gly Gln Val Leu Asp 1 5 aaa ttt caa cgt aag caa tgg aga caa aag caa atc cag aag ata aca 104 Lys Phe Gln Arg Lys Gln Trp Arg Gln Lys Gln Ile Gln Lys Ile Thr 10 15 20 gat aag gta ttt gat cgt gtc aaa agt ccg acc gga aat ggc act ctt 152 Asp Lys Val Phe Asp Arg Val Lys Ser Pro Thr Gly Asn Gly Thr Leu 25 30 35 aca ttt gaa gag ctg tat ata gct acc ctg att gtc tac aat gat ata 200 Thr Phe Glu Glu Leu Tyr Ile Ala Thr Leu Ile Val Tyr Asn Asp Ile 40 45 50 aac aag tat ttg ccg ggg ccg cac ttt gat cct cca tcg aaa gac aaa 248 Asn Lys Tyr Leu Pro Gly Pro His Phe Asp Pro Pro Ser Lys Asp Lys 55 60 65 70 atc aga gcc ttg atg cag gaa tgc gat atg gat gtc gat gga gaa ctt 296 Ile Arg Ala Leu Met Gln Glu Cys Asp Met Asp Val Asp Gly Glu Leu 75 80 85 aac cgt gag gaa ttt gtg aag ttc atg cag aag gtg aca gcc gat aca 344 Asn Arg Glu Glu Phe Val Lys Phe Met Gln Lys Val Thr Ala Asp Thr 90 95 100 ttc tct acg gtc agc cag gga ctg att atc tct ctg att ctg gcg cca 392 Phe Ser Thr Val Ser Gln Gly Leu Ile Ile Ser Leu Ile Leu Ala Pro 105 110 115 aca gtt gca ttg gcg acg aag agg gca aca gaa ggt gtt cca ggt gtg 440 Thr Val Ala Leu Ala Thr Lys Arg Ala Thr Glu Gly Val Pro Gly Val 120 125 130 ggg aaa gtg gtg caa aag gtg cct act tca att tat gca tcc ctg gtg 488 Gly Lys Val Val Gln Lys Val Pro Thr Ser Ile Tyr Ala Ser Leu Val 135 140 145 150 acc ctt gtt gtc gtt gca atc caa act gct agc gag gga tgc 530 Thr Leu Val Val Val Ala Ile Gln Thr Ala Ser Glu Gly Cys 155 160 tgattagagg ctttagttac ttgttcatga tacagaagga acagtcttgg tcaatttatt 590 tcttttttaa taggacataa ggaagttgta tatctttctt ctttcttcta ccaggttttg 650 ggggaagttg gaaagaacat acaaatgatt tcaactgcgt attggctgat cctcccattt 710 attaaaactt gtcgtgtcta gcatgagcga ttcaatattt gcaatatgca atatttgtaa 770 tgatgtctac attcagtgat tagtgtgatt gtgcagtttg ttgggaaaaa aaaaaaaaaa 830 aaaaaaaaaa aaaaaa 846 <210> 16 <211> 164 <212> PRT <213> Mesembryanthemum crystallinum <400> 16 Met Gly Gln Val Leu Asp Lys Phe Gln Arg Lys Gln Trp Arg Gln Lys 1 5 10 15 Gln Ile Gln Lys Ile Thr Asp Lys Val Phe Asp Arg Val Lys Ser Pro 20 25 30 Thr Gly Asn Gly Thr Leu Thr Phe Glu Glu Leu Tyr Ile Ala Thr Leu 35 40 45 Ile Val Tyr Asn Asp Ile Asn Lys Tyr Leu Pro Gly Pro His Phe Asp 50 55 60 Pro Pro Ser Lys Asp Lys Ile Arg Ala Leu Met Gln Glu Cys Asp Met 65 70 75 80 Asp Val Asp Gly Glu Leu Asn Arg Glu Glu Phe Val Lys Phe Met Gln 85 90 95 Lys Val Thr Ala Asp Thr Phe Ser Thr Val Ser Gln Gly Leu Ile Ile 100 105 110 Ser Leu Ile Leu Ala Pro Thr Val Ala Leu Ala Thr Lys Arg Ala Thr 115 120 125 Glu Gly Val Pro Gly Val Gly Lys Val Val Gln Lys Val Pro Thr Ser 130 135 140 Ile Tyr Ala Ser Leu Val Thr Leu Val Val Val Ala Ile Gln Thr Ala 145 150 155 160 Ser Glu Gly Cys  <210> 17 <211> 872 <212> DNA <213> Mesembryanthemum crystallinum <220> <221> CDS <222> (183) .. (569) <400> 17 aacaaaatgt ctctctcttt ctctttctct ttctctttct ctctcttcgt gggttgattg 60 agtaagctct gtccttttgc tctctgttga atgtactatc ttctgtgaac caaaggccaa 120 agattaacta ttggagattt ctctactcga aatttgtttt taggtgttga ccctgttgag 180 ct atg gcg aac aag ccc caa att cca acg aag aat tcg gcc ctc att 227 Met Ala Asn Lys Pro Gln Ile Pro Thr Lys Asn Ser Ala Leu Ile 1 5 10 15 gct att atc gcg gat gag gat act gta act gga ttt ttg ctg gct gga 275 Ala Ile Ile Ala Asp Glu Asp Thr Val Thr Gly Phe Leu Leu Ala Gly 20 25 30 gtt ggt aat gtt gat cta cga aga cag aca aat tac att att gtg gac 323 Val Gly Asn Val Asp Leu Arg Arg Gln Thr Asn Tyr Ile Ile Val Asp 35 40 45 aat aaa aca acg atg aag caa atc gaa gat gca ttc aag gag ttc aca 371 Asn Lys Thr Thr Met Lys Gln Ile Glu Asp Ala Phe Lys Glu Phe Thr 50 55 60 gca aga gag gac att gcg gtt gta cta atc agc caa tat gtt gca aat 419 Ala Arg Glu Asp Ile Ala Val Val Leu Ile Ser Gln Tyr Val Ala Asn 65 70 75 atg ata aga gta ttg gtt gat agc tac aac aaa cca atc ccg gca att 467 Met Ile Arg Val Leu Val Asp Ser Tyr Asn Lys Pro Ile Pro Ala Ile 80 85 90 95 ttg gag att cct tca aag gac cat cct tat gat cct aac cat gat tca 515 Leu Glu Ile Pro Ser Lys Asp His Pro Tyr Asp Pro Asn His Asp Ser 100 105 110 gtc ctt tca agg gtt aaa tac ctg ttc tct tct gaa tcg gca tca agc 563 Val Leu Ser Arg Val Lys Tyr Leu Phe Ser Ser Glu Ser Ala Ser Ser 115 120 125 aga ttt tagccatatg ctttgtaaag ttccctgctc ctgaatgttt ggtgattatg 619 Arg Phe agtttaaact agaaccagtc acattctgac ttggtatttt gaggcactgt ttgttttatg 679 ttcttaaaat aaggagtgta attacgactc catgaatcgg gatatgactc catgaatcgc 739 atgtatttct ttccatctca tttgaaagag tcgagcagcc atatcattta gtttcttcct 799 cttgcgaatg agcttggaag aaatgttttg gctataaaag atttcaactc ttggtacaaa 859 aaaaaaaaaa aaa 872 <210> 18 <211> 129 <212> PRT <213> Mesembryanthemum crystallinum <400> 18 Met Ala Asn Lys Pro Gln Ile Pro Thr Lys Asn Ser Ala Leu Ile Ala 1 5 10 15 Ile Ile Ala Asp Glu Asp Thr Val Thr Gly Phe Leu Leu Ala Gly Val 20 25 30 Gly Asn Val Asp Leu Arg Arg Gln Thr Asn Tyr Ile Ile Val Asp Asn 35 40 45 Lys Thr Thr Met Lys Gln Ile Glu Asp Ala Phe Lys Glu Phe Thr Ala 50 55 60 Arg Glu Asp Ile Ala Val Val Leu Ile Ser Gln Tyr Val Ala Asn Met 65 70 75 80 Ile Arg Val Leu Val Asp Ser Tyr Asn Lys Pro Ile Pro Ala Ile Leu 85 90 95 Glu Ile Pro Ser Lys Asp His Pro Tyr Asp Pro Asn His Asp Ser Val 100 105 110 Leu Ser Arg Val Lys Tyr Leu Phe Ser Ser Glu Ser Ala Ser Ser Arg 115 120 125 Phe  <210> 19 <211> 647 <212> DNA <213> Mesembryanthemum crystallinum <220> <221> CDS <222> (64) .. (426) <400> 19 cttgtttttc tctctcctct ctctctctct tctccgcacc ctcaggcagt gaaggtagca 60 aca atg gcg tac gcg atg aag cca acg aag ccc ggg atg gag gaa tcc 108 Met Ala Tyr Ala Met Lys Pro Thr Lys Pro Gly Met Glu Glu Ser 1 5 10 15 cag gag cag att cac aag atc agg atc act ctt tct tct aag aac gtc 156 Gln Glu Gln Ile His Lys Ile Arg Ile Thr Leu Ser Ser Lys Asn Val 20 25 30 aag aac ctt gag aaa gtg tgt gct gat ctt gta cgc ggt gca aag gac 204 Lys Asn Leu Glu Lys Val Cys Ala Asp Leu Val Arg Gly Ala Lys Asp 35 40 45 aag cgc ctc agg gtt aag gga cca gtg agg atg ccc acc aag gtt ctg 252 Lys Arg Leu Arg Val Lys Gly Pro Val Arg Met Pro Thr Lys Val Leu 50 55 60 aag atc aca aca agg aag tct ccc tgt ggt gaa gga acc aac acc ttt 300 Lys Ile Thr Thr Arg Lys Ser Pro Cys Gly Glu Gly Thr Asn Thr Phe 65 70 75 gac aga ttt gag ttg cgt gtt cac aag aga gtc att gac ctc ttc agc 348 Asp Arg Phe Glu Leu Arg Val His Lys Arg Val Ile Asp Leu Phe Ser 80 85 90 95 tcc cca gac gtg gtc aag cag atc acc tcc atc acc att gaa cct ggt 396 Ser Pro Asp Val Val Lys Gln Ile Thr Ser Ile Thr Ile Glu Pro Gly 100 105 110 gtt gag gtt gag gtt aca ata gct gac tct tagacatgcc tgttgaagtt 446 Val Glu Val Glu Val Thr Ile Ala Asp Ser 115 120 gtcgtcgttg tagggctgtt gtagctgtct catatagtgg tgctatctca ctaagaattt 506 tgaagatact aaattgtttg tttgaaagag atgttttctt tagctgtaat gttatgtttt 566 tgaaggtgtt ggaacatgca ttatttgtta atgctttatc aatagaactt ccaatttgaa 626 tgcaaaaaaa aaaaaaaaaa a 647 <210> 20 <211> 121 <212> PRT <213> Mesembryanthemum crystallinum <400> 20 Met Ala Tyr Ala Met Lys Pro Thr Lys Pro Gly Met Glu Glu Ser Gln 1 5 10 15 Glu Gln Ile His Lys Ile Arg Ile Thr Leu Ser Ser Lys Asn Val Lys 20 25 30 Asn Leu Glu Lys Val Cys Ala Asp Leu Val Arg Gly Ala Lys Asp Lys 35 40 45 Arg Leu Arg Val Lys Gly Pro Val Arg Met Pro Thr Lys Val Leu Lys 50 55 60 Ile Thr Thr Arg Lys Ser Pro Cys Gly Glu Gly Thr Asn Thr Phe Asp 65 70 75 80 Arg Phe Glu Leu Arg Val His Lys Arg Val Ile Asp Leu Phe Ser Ser 85 90 95 Pro Asp Val Val Lys Gln Ile Thr Ser Ile Thr Ile Glu Pro Gly Val 100 105 110 Glu Val Glu Val Thr Ile Ala Asp Ser 115 120 <210> 21 <211> 686 <212> DNA <213> Sueada japonica <220> <221> CDS <222> (62) .. (493) <400> 21 acaccattca caaaacacat taaaaaaaaa cactacttct ttctttctta gccacttgaa 60 a atg gcc tac tca aag gct gta ctc ctt gcc ctt atc ttt gct gtg act 109 Met Ala Tyr Ser Lys Ala Val Leu Leu Ala Leu Ile Phe Ala Val Thr 1 5 10 15 ctt gtc att gcc tct cag gtc tca gct cgt gaa ctt gct gag gag aca 157 Leu Val Ile Ala Ser Gln Val Ser Ala Arg Glu Leu Ala Glu Glu Thr 20 25 30 caa tct gtg gag gag tct aag gga tac ggt ggt ggg cac gga ggt cac 205 Gln Ser Val Glu Glu Ser Lys Gly Tyr Gly Gly Gly His Gly Gly His 35 40 45 tat ggt ggt ggt cac tat ggt ggt gga cac aga cac ggt ggc cat gga 253 Tyr Gly Gly Gly His Tyr Gly Gly Gly His Arg His Gly Gly His Gly 50 55 60 cac tac gca act gag gaa gca gag aac aag aat gaa gcc gta gaa cct 301 His Tyr Ala Thr Glu Glu Ala Glu Asn Lys Asn Glu Ala Val Glu Pro 65 70 75 80 caa ggc ggc tat ggt cac gga cac gga gga ggc tac gga cac ggt ggt 349 Gln Gly Gly Tyr Gly His Gly His Gly Gly Gly Tyr Gly His Gly Gly 85 90 95 ggc tac gga cac ggt gga ggc tac gga cac gga ggt ggc tac ggg cac 397 Gly Tyr Gly His Gly Gly Gly Tyr Gly His Gly Gly Gly Tly Gyr His 100 105 110 ggt ggt ggc tac gga cat gga ggt ggt tat gga cac ggt gga cac ggt 445 Gly Gly Gly Tyr Gly His Gly Gly Gly Tyr Gly His Gly Gly His Gly 115 120 125 gga cat ggt ggt cat ggt cac tac gcc aag act acc gag gaa caa aat 493 Gly His Gly Gly His Gly His Tyr Ala Lys Thr Thr Glu Glu Gln Asn 130 135 140 taagttatgg gttactaaaa cttaaattgt acgttgtcaa ataaaatgta ctttatgatt 553 ttacatgagt atgcatgtaa ttcatcataa gcttcaagga ctatcttgta ctctatgtta 613 tatacctata tgaaatggaa gcgtgacttt tattactgta aaaaaaaaaa aaaaaaaaaa 673 aaaaaaaaaa aaa 686 <210> 22 <211> 144 <212> PRT <213> Sueada japonica <400> 22 Met Ala Tyr Ser Lys Ala Val Leu Leu Ala Leu Ile Phe Ala Val Thr 1 5 10 15 Leu Val Ile Ala Ser Gln Val Ser Ala Arg Glu Leu Ala Glu Glu Thr 20 25 30 Gln Ser Val Glu Glu Ser Lys Gly Tyr Gly Gly Gly His Gly Gly His 35 40 45 Tyr Gly Gly Gly His Tyr Gly Gly Gly His Arg His Gly Gly His Gly 50 55 60 His Tyr Ala Thr Glu Glu Ala Glu Asn Lys Asn Glu Ala Val Glu Pro 65 70 75 80 Gln Gly Gly Tyr Gly His Gly His Gly Gly Gly Tyr Gly His Gly Gly 85 90 95 Gly Tyr Gly His Gly Gly Gly Tyr Gly His Gly Gly Gly Tly Gyr His 100 105 110 Gly Gly Gly Tyr Gly His Gly Gly Gly Tyr Gly His Gly Gly His Gly 115 120 125 Gly His Gly Gly His Gly His Tyr Ala Lys Thr Thr Glu Glu Gln Asn 130 135 140 <210> 23 <211> 683 <212> DNA <213> Salsola komarovii <220> <221> CDS <222> (48) .. (362) <400> 23 gttaagatat tatattgcaa ctttacaaag catttctgca actaaat atg gcc ttt 56 Met Ala Phe 1 tcc aaa cct cta att gct tct cta ctt ctt tct ctc ttt gtt ctt cag 104 Ser Lys Pro Leu Ile Ala Ser Leu Leu Leu Ser Leu Phe Val Leu Gln 5 10 15 ttt gtt cat gca gtt gaa cct att tca tcc tcc aat caa gtg ggt agc 152 Phe Val His Ala Val Glu Pro Ile Ser Ser Ser Asn Gln Val Gly Ser 20 25 30 35 aac act gga ggt acc tca gag agt aaa gtg gat tgt ggg gcg gca tgt 200 Asn Thr Gly Gly Thr Ser Glu Ser Lys Val Asp Cys Gly Ala Ala Cys 40 45 50 acg gtg agg tgc agc gcc tcg aag agg cca aac cta tgc aac agg tca 248 Thr Val Arg Cys Ser Ala Ser Lys Arg Pro Asn Leu Cys Asn Arg Ser 55 60 65 tgt ggc agt tgt tgc aag acg tgc aac tgc gtg cca cca ggc act tcc 296 Cys Gly Ser Cys Cys Lys Thr Cys Asn Cys Val Pro Pro Gly Thr Ser 70 75 80 ggc aac tac gaa gcc tgc cct tgt tac gcc aac ttg acc acc cac ggc 344 Gly Asn Tyr Glu Ala Cys Pro Cys Tyr Ala Asn Leu Thr Thr His Gly 85 90 95 aat cga cac aag tgc cct taattaacaa gaattgttta gttgtttatt 392 Asn Arg His Lys Cys Pro 100 105 acatccgtac catgtaacgt actcctattt acactactag agtactagta ataaacattt 452 ttaggcacgg tccagttgtt catgtagcta gtggtatatt gagtcataaa tgagtgattg 512 aaaatgagat atgataaaag tgtattatct acattgtagt actgttttgt atcatagtgt 572 agtgatgttt atttttcgta cctttaattt gttactttgt attccctttc attctatcta 632 tttacaatcc ttttgtaagt ttatgtgaaa aaaaaaaaaa aaaaaaaaaa a 683 <210> 24 <211> 105 <212> PRT <213> Salsola komarovii <400> 24 Met Ala Phe Ser Lys Pro Leu Ile Ala Ser Leu Leu Leu Ser Leu Phe 1 5 10 15 Val Leu Gln Phe Val His Ala Val Glu Pro Ile Ser Ser Ser Asn Gln 20 25 30 Val Gly Ser Asn Thr Gly Gly Thr Ser Glu Ser Lys Val Asp Cys Gly 35 40 45 Ala Ala Cys Thr Val Arg Cys Ser Ala Ser Lys Arg Pro Asn Leu Cys 50 55 60 Asn Arg Ser Cys Gly Ser Cys Cys Lys Thr Cys Asn Cys Val Pro Pro 65 70 75 80 Gly Thr Ser Gly Asn Tyr Glu Ala Cys Pro Cys Tyr Ala Asn Leu Thr 85 90 95 Thr His Gly Asn Arg His Lys Cys Pro 100 105 <210> 25 <211> 803 <212> DNA <213> Salsola komarovii <220> <221> CDS <222> (51) .. (593) <400> 25 cgcagacgct tcagctcttt ctctctcttt ctctctcctc accgtgaaag atg ggg 56 Met Gly 1 ttg tca ttt acc aaa ttg ttt agc cgg ttg ttc gct aag aag gaa atg 104 Leu Ser Phe Thr Lys Leu Phe Ser Arg
Leu Phe Ala Lys Lys Glu Met 5 10
Fifteen cgt atc ctt atg gtc ggt ctc gat gcc
gct ggt aaa acc acc att ctc 152 Arg Ile Leu Met Val Gly Leu Asp Ala
Ala Gly Lys Thr Thr Ile Leu 20 25
30 tat aaa ctc aag ctg gga gag att gtc
acc acc att cct acc att gga 200 Tyr Lys Leu Lys Leu Gly Glu Ile Val
Thr Thr Ile Pro Thr Ile Gly 35 40
45 50 ttt aat gtg gag act gta gaa tac aag
aac atc agc ttc act gtg tgg 248 Phe Asn Val Glu Tyr Val Glu Tyr Lys
Asn Ile Ser Phe Thr Val Trp 55
60 65 gat gtc ggg ggt ca gac aag att cgt
cca tttg tgg aga cat tact tttc 296 Asp Val Gly Gly Gly Asn Asp Lys Ile Arg
Pro Leu Trp Arg His Tyr Phe 70 75
80 caa aac acc caa ggt ctc atc ttt ggtg
gtt gac agt aat ga t cgt gac 344 Gln Asn Thr Gln Gly Leu Ile Phe Val
Val Asp Ser Asn Asp Arg Asp 85 90
95 cgt gtc gtt gag gca aga ga t gaa ctg
cat agg atg tta aat gag gat 392 Arg Val Val Glu Ala Arg Asp Glu Leu
His Arg Met Leu Asn Glu Asp 100 105
110 ga tta cga gat gca gtg ttg ttg gtg
ttt gca aac aag caa gat ctt 440 115 120 125 130 ccc aat gca atg aat gct gct gag atc act gat aag ctt ggt ctc cat 488 Pro Asn Ala Met Asn Ala Ala Glu Ile Thr Asp Lys Leu Gly Leu His 135 140 145 tct cta cgt caa cgc cat tgg tac ata caa agc aca tgt gcc acc tct 536 Ser Leu Arg Gln Arg His Trp Tyr Ile Gln Ser Thr Cys Ala Thr Ser 150 155 160 gga gaa ggg ctt tac gag ggt ctg gac tgg ctc tca aac aat atc gct 584 Gly Glu Gly Leu Tyr Glu Gly Leu Asp Trp Leu Ser Asn Asn Ile Ala 165 170 175 agc aag gct taaaagtaac agaacgagta aggttagctt tctcagagaa 633 Ser Lys Ala 180 gaagctggag tataggctga ggactatcgt tactgctagt gttacccttt ttatttttgc 693 catttatatg ttcacatttt tggttcctat cggacaagaa ttattttctg cgtttatgtt 753 gacttgttat aataccatac tttttagttg aaaaaaaaaa aaaaaaaaaa 803 <210> 26 <211> 181 <212> PRT <213> Salsola komarovii <400> 26 Met Gly Leu Ser Phe Thr Lys Leu Phe Ser Arg Leu Phe Ala Lys Lys 1 5 10 15 Glu Met Arg Ile Leu Met Val Gly Leu Asp Ala Ala Gly Lys Thr Thr 20 25 30 Ile Leu Tyr Lys Leu Lys Leu Gly Glu Ile Val Thr Thr Ile Pro Thr 35 40 45 Ile Gly Phe Asn Val Glu Thr Val Glu Tyr Lys Asn Ile Ser Phe Thr 50 55 60 Val Trp Asp Val Gly Gly Gln Asp Lys Ile Arg Pro Leu Trp Arg His 65 70 75 80 Tyr Phe Gln Asn Thr Gln Gly Leu Ile Phe Val Val Asp Ser Asn Asp 85 90 95 Arg Asp Arg Val Val Glu Ala Arg Asp Glu Leu His Arg Met Leu Asn 100 105 110 Glu Asp Glu Leu Arg Asp Ala Val Leu Leu Val Phe Ala Asn Lys Gln 115 120 125 Asp Leu Pro Asn Ala Met Asn Ala Ala Glu Ile Thr Asp Lys Leu Gly 130 135 140 Leu His Ser Leu Arg Gln Arg His Trp Tyr Ile Gln Ser Thr Cys Ala 145 150 155 160 Thr Ser Gly Glu Gly Leu Tyr Glu Gly Leu Asp Trp Leu Ser Asn Asn 165 170 175 Ile Ala Ser Lys Ala 180 <210> 27 <211> 680 <212> DNA <213> Avicennia marina <220> <221> CDS <222> (161) .. (454) <400> 27 ctaaaagcca aaggcaagat aagaaacagg ttcctttagc tatcttcctc gtctcgctgc 60 tgcaaaagtt ccatccccag aagatcagga aaacccttct gcagcagcac tctaataatc 120 ctccaatttt gattcaagag aagaaacaaa ataaacagaa atg gct cgc tct ttc 175 Met Ala Arg Ser Phe 1 5 tcc aac gct aag acc gtc tct gct gtc att gcc aac gaa atc tca gct 223 Ser Asn Ala Lys Thr Val Ser Ala Val Ile Ala Asn Glu Ile Ser Ala 10 15 20 ctt gtc acc agg agg ggt tat gct gct ctc gca cag ggc gtt gtt tcg 271 Leu Val Thr Arg Arg Gly Tyr Ala Ala Leu Ala Gln Gly Val Val Ser 25 30 35 agc agc gcg aga agc ggc ggc gct ccg aac gtg atg ctg aag aaa gga 319 Ser Ser Ala Arg Ser Gly Gly Ala Pro Asn Val Met Leu Lys Lys Gly 40 45 50 tcc gaa gaa tcc ggg aag aca gca tgg gtg ccc gac ccg gac acc ggc 367 Ser Glu Glu Ser Gly Lys Thr Ala Trp Val Pro Asp Pro Asp Thr Gly 55 60 65 tac tac cga ccg gga aac gag gac aag gcc gcg ctg gac ccg gtc gag 415 Tyr Tyr Arg Pro Gly Asn Glu Asp Lys Ala Ala Leu Asp Pro Val Glu 70 75 80 85 ctg cgg gag atg ctc atc aag aac aag ccc agc cga caa tgaatgaacc 464 Leu Arg Glu Met Leu Ile Lys Asn Lys Pro Ser Arg Gln 90 95 aagaattgtg ggattctcat taattcctcc cctgttctgg tccatcgtcg gaatctgaac 524 ctgttgttcg tctagaaatt cgttcccatg gaaatctatc aaagtctgta ttcttgccat 584 ggctcttcct gtcccatata tgtatgtcct caggtgtggc ctggggtggt ttgatagata 644 tataaaatgt ggtgaattta aaaaaaaaaa aaaaaa 680 <210> 28 <211> 98 <212> PRT <213> Avicennia marina <400> 28 Met Ala Arg Ser Phe Ser Asn Ala Lys Thr Val Ser Ala Val Ile Ala 1 5 10 15 Asn Glu Ile Ser Ala Leu Val Thr Arg Arg Gly Tyr Ala Ala Leu Ala 20 25 30 Gln Gly Val Val Ser Ser Ser Ala Arg Ser Gly Gly Ala Pro Asn Val 35 40 45 Met Leu Lys Lys Gly Ser Glu Glu Ser Gly Lys Thr Ala Trp Val Pro 50 55 60 Asp Pro Asp Thr Gly Tyr Tyr Arg Pro Gly Asn Glu Asp Lys Ala Ala 65 70 75 80 Leu Asp Pro Val Glu Leu Arg Glu Met Leu Ile Lys Asn Lys Pro Ser 85 90 95 Arg Gln <210> 29 <211> 490 <212> DNA <213> Avicennia marina <220> <221> CDS <222> (20) .. (349) <400> 29 tcggctgggc aaagaaggg atg gcg att cca tcg gaa att cgg gac ttt att 52 Met Ala Ile Pro Ser Glu Ile Arg Asp Phe Ile 1 5 10 gct agc cgc aac aga tct ttg gtg atc gca tct cca aag gaa gat gag 100 Ala Ser Arg Asn Arg Ser Leu Val Ile Ala Ser Pro Lys Glu Asp Glu 15 20 25 aaa att ctc cgc tca agg cag tgc acc gaa gaa ggg gcg cgt gca gga 148 Lys Ile Leu Arg Ser Arg Gln Cys Thr Glu Glu Gly Ala Arg Ala Gly 30 35 40 gcc aaa gct gct gca gtt gct tgc gtt gcc agc gcc att ccc act ctg 196 Ala Lys Ala Ala Ala Val Ala Cys Val Ala Ser Ala Ile Pro Thr Leu 45 50 55 gta gct gtt cga acg att ccg tgg gca aag gca aac ctc aac tat aca 244 Val Ala Val Arg Thr Ile Pro Trp Ala Lys Ala Asn Leu Asn Tyr Thr 60 65 70 75 gcc cag gca ctc att ata tct tct gca tcc ata gcg gca tac ttt atc 292 Ala Gln Ala Leu Ile Ile Ser Ser Ala Ser Ile Ala Ala Tyr Phe Ile 80 85 90 gct gct gac aaa acc atc tta gag tgc gca cgg aaa aat gca gag tac 340 Ala Ala Asp Lys Thr Ile Leu Glu Cys Ala Arg Lys Asn Ala Glu Tyr 95 100 105 aaa tcg gct taagatgatg tgtaagacaa tgtgctcagc ttgcaatgct 389 Lys Ser Ala 110 tgccatgact tgtgtttatg tgtatttcaa gtttctgaaa ctagcatttt gattttgtgt 449 tccaatgcaa tgagcattat ggaaaaaaaa aaaaaaaaaa a 490 <210> 30 <211> 110 <212> PRT <213> Avicennia marina <400> 30 Met Ala Ile Pro Ser Glu Ile Arg Asp Phe Ile Ala Ser Arg Asn Arg 1 5 10 15 Ser Leu Val Ile Ala Ser Pro Lys Glu Asp Glu Lys Ile Leu Arg Ser 20 25 30 Arg Gln Cys Thr Glu Glu Gly Ala Arg Ala Gly Ala Lys Ala Ala Ala 35 40 45 Val Ala Cys Val Ala Ser Ala Ile Pro Thr Leu Val Ala Val Arg Thr 50 55 60 Ile Pro Trp Ala Lys Ala Asn Leu Asn Tyr Thr Ala Gln Ala Leu Ile 65 70 75 80 Ile Ser Ser Ala Ser Ile Ala Ala Tyr Phe Ile Ala Ala Asp Lys Thr 85 90 95 Ile Leu Glu Cys Ala Arg Lys Asn Ala Glu Tyr Lys Ser Ala 100 105 110 <210> 31 <211> 592 <212> DNA <213> Avicennia marina <220> <221> CDS <222> (75) .. (320) <400> 31 gcagtctcag ccttcctgct ctcctggtgc cttcaaattt gtgaatttct cgagtgctaa 60 aagattcagc caag atg cag aac gaa gag ggg caa aac atg gat ctc tac 110 Met Gln Asn Glu Glu Gly Gln Asn Met Asp Leu Tyr 1 5 10 atc ccc agg aaa tgc tct gcc acg aac agg ctg atc acc tcc aag gat 158 Ile Pro Arg Lys Cys Ser Ala Thr Asn Arg Leu Ile Thr Ser Lys Asp 15 20 25 cat gct tct gtc cag atc aat gtt ggg cac ttg gat gag aat ggc cga 206 His Ala Ser Val Gln Ile Asn Val Gly His Leu Asp Glu Asn Gly Arg 30 35 40 tac act ggc caa tac tct acc ttt gct ctt tgt gga ttc atc cgt gct 254 Tyr Thr Gly Gln Tyr Ser Thr Phe Ala Leu Cys Gly Phe Ile Arg Ala 45 50 55 60 cag ggt gat gct gac agt gct ctt gat agg ctc tgg cag aaa aag aaa 302 Gln Gly Asp Ala Asp Ser Ala Leu Asp Arg Leu Trp Gln Lys Lys Lys 65 70 75 gtc gaa acc agg cag cag tgatcctgct caattcagca gtgaaagttt 350 Val Glu Thr Arg Gln Gln 80 tttgggtttt gttctgtgtt gtgttattta tgcttttcca gaatcaattt ctgtactgga 410 ttgagtatta aaaatgtgga gctaaaggtt gggagacctg atgcctttgt tactcgagta 470 atcacaagta gatactgggc ttgtaatagc gtgataattg tgccttgctc ttgcctcatt 530 gactacgaat cagttatgtg attagacaat gttaatctcc aaaaaaaaaa aaaaaaaaaa 590 aa 592 <210> 32 <211> 82 <212> PRT <213> Avicennia marina <400> 32 Met Gln Asn Glu Glu Gly Gln Asn Met Asp Leu Tyr Ile Pro Arg Lys 1 5 10 15 Cys Ser Ala Thr Asn Arg Leu Ile Thr Ser Lys Asp His Ala Ser Val 20 25 30 Gln Ile Asn Val Gly His Leu Asp Glu Asn Gly Arg Tyr Thr Gly Gln 35 40 45 Tyr Ser Thr Phe Ala Leu Cys Gly Phe Ile Arg Ala Gln Gly Asp Ala 50 55 60 Asp Ser Ala Leu Asp Arg Leu Trp Gln Lys Lys Lys Val Glu Thr Arg 65 70 75 80 Gln Gln  <210> 33 <211> 1806 <212> DNA <213> Avicennia marina <220> <221> CDS <222> (362) .. (1552) <400> 33 tgtgaaggta aagtctacag catatttcgc gccgctcgtt tgattacgtg ttgcttttat 60 ttgggaattt gatagcgctg agtagccgat gccgctggag ggtattgttg attttaggaa 120 tacgggtttg tttgattcgc agttttactg tctctagggt tgggccctga ggcttctggg 180 atttgggatt taatcgctga tcgaacagtt tcctggagaa aatactccta gtgcgcatat 240 atctgatttg ctgacgagaa attgatacac ggttatgcga ttgagttttg tttgcgccaa 300 agatactccg agtgctcgct agatgtggat aatccggagg gctgtttcga tgagatgagg 360 g atg tta tca ggg tta atg aac ttc ctg tgg gcc tgt ttt cgg cca agg 409 Met Leu Ser Gly Leu Met Asn Phe Leu Trp Ala Cys Phe Arg Pro Arg 1 5 10 15 gcg gat cga agt gtt cac acg ggt tca gat gca ggc ggt cgt cag gat 457 Ala Asp Arg Ser Val His Thr Gly Ser Asp Ala Gly Gly Arg Gln Asp 20 25 30 ggg ctt tta tgg tat aag gac ttg ggg caa cat atc aat gga gag ttt 505 Gly Leu Leu Trp Tyr Lys Asp Leu Gly Gln His Ile Asn Gly Glu Phe 35 40 45 tca atg gct gta gtt caa gca aat aac tta cta gag gat cag agt caa 553 Ser Met Ala Val Val Gln Ala Asn Asn Leu Leu Glu Asp Gln Ser Gln 50 55 60 ctt gaa tct ggt tgc ctg agc ttg agt gat tca gga caa tat ggc act 601 Leu Glu Ser Gly Cys Leu Ser Leu Ser Asp Ser Gly Gln Tyr Gly Thr 65 70 75 80 ttt gtg ggg att tat gat gga cat gga ggt cct gag acc tct cgg ttt 649 Phe Val Gly Ile Tyr Asp Gly His Gly Gly Pro Glu Thr Ser Arg Phe 85 90 95 atc aat gac cat ctc ttc caa cat ata aag aga ttc aca gct gag cat 697 Ile Asn Asp His Leu Phe Gln His Ile Lys Arg Phe Thr Ala Glu His 100 105 110 caa tca atg tca gct gag gtc att cac aag gcc att caa gcg act gaa 745 Gln Ser Met Ser Ala Glu Val Ile His Lys Ala Ile Gln Ala Thr Glu 115 120 125 gaa ggt ttt ttc tcg gtt gtt agc aga caa tgg tcc atg caa cca cag 793 Glu Gly Phe Phe Ser Val Val Ser Arg Gln Trp Ser Met Gln Pro Gln 130 135 140 att gca gca gtt ggc tct tgc tgc ctt gtt ggt gtc atc tgt agt ggc 841 Ile Ala Ala Val Gly Ser Cys Cys Leu Val Gly Val Ile Cys Ser Gly 145 150 155 160 act ctt tat gtt tcc aac ctt ggt gat tcc cgt gct gtt ctt ggg acg 889 Thr Leu Tyr Val Ser Asn Leu Gly Asp Ser Arg Ala Val Leu Gly Thr 165 170 175 ctt tcc aag gct aca ggg gaa gta cag gct act caa ctc tca aca gag 937 Leu Ser Lys Ala Thr Gly Glu Val Gln Ala Thr Gln Leu Ser Thr Glu 180 185 190 cat aat gca agt ttt gag tct gtg aga cgg gaa ctg cag tct ctg cac 985 His Asn Ala Ser Phe Glu Ser Val Arg Arg Glu Leu Gln Ser Leu His 195 200 205 cca gat gac tca cag att gtg gtt cta aag cat aat gta tgg cga gtg 1033 Pro Asp Asp Ser Gln Ile Val Val Leu Lys His Asn Val Trp Arg Val 210 215 220 aag ggt ctt ata cag atc tca aga tca att gga gat gtg tat ttg aaa 1081 Lys Gly Leu Ile Gln Ile Ser Arg Ser Ile Gly Asp Val Tyr Leu Lys 225 230 235 240 aag gct gaa ttc aac agg gag cct cta tat cag aaa ttt cga ctt cgt 1129 Lys Ala Glu Phe Asn Arg Glu Pro Leu Tyr Gln Lys Phe Arg Leu Arg 245 250 255 gaa gct ttc aaa aga cca att ttg agc tca gaa cca gaa act act gtg 1177 Glu Ala Phe Lys Arg Pro Ile Leu Ser Ser Glu Pro Glu Thr Thr Val 260 265 270 cac cag ctg ctg cct cat gat caa ttc att atc ttc gca tca gat ggc 1225 His Gln Leu Leu Pro His Asp Gln Phe Ile Ile Phe Ala Ser Asp Gly 275 280 285 ctt tgg gag cac ctt tcc aac caa gaa gca gtt gat ctt gtt cag aaa 1273 Leu Trp Glu His Leu Ser Asn Gln Glu Ala Val Asp Leu Val Gln Lys 290 295 300 cat cca cac aat ggg att gct aga aga tta gta aaa gca gct ttg caa 1321 His Pro His Asn Gly Ile Ala Arg Arg Leu Val Lys Ala Ala Leu Gln 305 310 315 320 gag gca gca aag aaa agg gaa atg agg tac tcg gat ttg aag aaa att 1369 Glu Ala Ala Lys Lys Arg Glu Met Arg Tyr Ser Asp Leu Lys Lys Ile 325 330 335 gac cgt ggg gtt cgc cgt cat ttc cat gat gac atc act gtt gtg gtg 1417 Asp Arg Gly Val Arg Arg His Phe His Asp Asp Ile Thr Val Val Val 340 345 350 ggt ttt ctt gac tca cac ctt gtg agc cgg gct agc tca gtc cgg ggc 1465 Val Phe Leu Asp Ser His Leu Val Ser Arg Ala Ser Ser Val Arg Gly 355 360 365 cca aac atc tcc gtg aaa ggt ggc ggc atc agt ctg cct ccc aat gct 1513 Pro Asn Ile Ser Val Lys Gly Gly Gly Ile Ser Leu Pro Pro Asn Ala 370 375 380 ctt gca cct tgt gcc aca cca acg gag cca gtc cca aat tgatactgct 1562 Leu Ala Pro Cys Ala Thr Pro Thr Glu Pro Val Pro Asn 385 390 395 gtctcttcta atgttatttc ccgttagtcc tgttgtacta ttgttatgtg aatacaggta 1622 gcttcttaac ggataacagc ggcccttgaa ttctttaatc catactgtaa cttttaaccg 1682 gagactatta cttggcatag tttcaatgcc caagggatac atagactggg acaagccatc 1742 ttggcggtga caatcatcat agttaagttt tctgggcata tctttcaaaa aaaaaaaaaa 1802 aaaa 1806 <210> 34 <211> 397 <212> PRT <213> Avicennia marina <400> 34 Met Leu Ser Gly Leu Met Asn Phe Leu Trp Ala Cys Phe Arg Pro Arg 1 5 10 15 Ala Asp Arg Ser Val His Thr Gly Ser Asp Ala Gly Gly Arg Gln Asp 20 25 30 Gly Leu Leu Trp Tyr Lys Asp Leu Gly Gln His Ile Asn Gly Glu Phe 35 40 45 Ser Met Ala Val Val Gln Ala Asn Asn Leu Leu Glu Asp Gln Ser Gln 50 55 60 Leu Glu Ser Gly Cys Leu Ser Leu Ser Asp Ser Gly Gln Tyr Gly Thr 65 70 75 80 Phe Val Gly Ile Tyr Asp Gly His Gly Gly Pro Glu Thr Ser Arg Phe 85 90 95 Ile Asn Asp His Leu Phe Gln His Ile Lys Arg Phe Thr Ala Glu His 100 105 110 Gln Ser Met Ser Ala Glu Val Ile His Lys Ala Ile Gln Ala Thr Glu 115 120 125 Glu Gly Phe Phe Ser Val Val Ser Arg Gln Trp Ser Met Gln Pro Gln 130 135 140 Ile Ala Ala Val Gly Ser Cys Cys Leu Val Gly Val Ile Cys Ser Gly 145 150 155 160 Thr Leu Tyr Val Ser Asn Leu Gly Asp Ser Arg Ala Val Leu Gly Thr 165 170 175 Leu Ser Lys Ala Thr Gly Glu Val Gln Ala Thr Gln Leu Ser Thr Glu 180 185 190 His Asn Ala Ser Phe Glu Ser Val Arg Arg Glu Leu Gln Ser Leu His 195 200 205 Pro Asp Asp Ser Gln Ile Val Val Leu Lys His Asn Val Trp Arg Val 210 215 220 Lys Gly Leu Ile Gln Ile Ser Arg Ser Ile Gly Asp Val Tyr Leu Lys 225 230 235 240 Lys Ala Glu Phe Asn Arg Glu Pro Leu Tyr Gln Lys Phe Arg Leu Arg 245 250 255 Glu Ala Phe Lys Arg Pro Ile Leu Ser Ser Glu Pro Glu Thr Thr Val 260 265 270 His Gln Leu Leu Pro His Asp Gln Phe Ile Ile Phe Ala Ser Asp Gly 275 280 285 Leu Trp Glu His Leu Ser Asn Gln Glu Ala Val Asp Leu Val Gln Lys 290 295 300 His Pro His Asn Gly Ile Ala Arg Arg Leu Val Lys Ala Ala Leu Gln 305 310 315 320 Glu Ala Ala Lys Lys Arg Glu Met Arg Tyr Ser Asp Leu Lys Lys Ile 325 330 335 Asp Arg Gly Val Arg Arg His Phe His Asp Asp Ile Thr Val Val Val 340 345 350 Val Phe Leu Asp Ser His Leu Val Ser Arg Ala Ser Ser Val Arg Gly 355 360 365 Pro Asn Ile Ser Val Lys Gly Gly Gly Ile Ser Leu Pro Pro Asn Ala 370 375 380 Leu Ala Pro Cys Ala Thr Pro Thr Glu Pro Val Pro Asn 385 390 395 <210> 35 <211> 743 <212> DNA <213> Mesembryanthemum crystallinum <220> <221> CDS <222> (1) .. (420) <400> 35 cct gag cta gca cct aaa gat ggg gat ttc cgt ttc aat atc tct gag 48 Pro Glu Leu Ala Pro Lys Asp Gly Asp Phe Arg Phe Asn Ile Ser Glu 1 5 10 15 ctt gaa gct atg cta cca gct gga act gta gat cat gct gtt gaa agg 96 Leu Glu Ala Met Leu Pro Ala Gly Thr Val Asp His Ala Val Glu Arg 20 25 30 att tat caa gag atg ccg cgg tgg gaa gag act gtt tta ggt tcc agg 144 Ile Tyr Gln Glu Met Pro Arg Trp Glu Glu Thr Val Leu Gly Ser Arg 35 40 45 agc aga tat gag cat gtc att cag gca ctt gca gat aaa tac cct tca 192 Ser Arg Tyr Glu His Val Ile Gln Ala Leu Ala Asp Lys Tyr Pro Ser 50 55 60 gaa aat ttg ttg cta gtt acg cat ggt gaa ggt gtt ggg act tca gtt 240 Glu Asn Leu Leu Leu Val Thr His Gly Glu Gly Val Gly Thr Ser Val 65 70 75 80 gca acg ttt ttg aaa ggc gct gtt gtt tat gaa gta aag tat tgt gct 288 Ala Thr Phe Leu Lys Gly Ala Val Val Tyr Glu Val Lys Tyr Cys Ala 85 90 95 tat tca caa gca aca aga cgc atc agc tat gga gaa ggc gag tca ttt 336 Tyr Ser Gln Ala Thr Arg Arg Ile Ser Tyr Gly Glu Gly Glu Ser Phe 100 105 110 act gct ggt acc ttt cag ttg gtc act gcc tca gac caa acc ggt att 384 Thr Ala Gly Thr Phe Gln Leu Val Thr Ala Ser Asp Gln Thr Gly Ile 115 120 125 ggt tac tac aca tct agc agc ttg tct gat ggt gta tgacttatcg 430 Gly Tyr Tyr Thr Ser Ser Ser Leu Ser Asp Gly Val 130 135 140 gaactcccga gtttctgcat tctgaaaggt gctttttgat ttccgaataa ttcttcaaat 490 ccacatgtca gaagatccat tctttaggtc agatgtctat ctactgctcc cagccttgag 550 ctgctcatgg gtattggtgc ccttctattt ttaggtagag tctttgagta agccttgcca 610 catcaaggcc tcagattatt gaatgtacaa cagaataggt tgtagcttca ttggctagta 670 cagtgacctc tttcatgggt ctgaaacatc aatataaagg tttgaatggc aaaaaaaaaa 730 aaaaaaaaaa aaa 743 <210> 36 <211> 140 <212> PRT <213> Mesembryanthemum crystallinum <400> 36 Pro Glu Leu Ala Pro Lys Asp Gly Asp Phe Arg Phe Asn Ile Ser Glu 1 5 10 15 Leu Glu Ala Met Leu Pro Ala Gly Thr Val Asp His Ala Val Glu Arg 20 25 30 Ile Tyr Gln Glu Met Pro Arg Trp Glu Glu Thr Val Leu Gly Ser Arg 35 40 45 Ser Arg Tyr Glu His Val Ile Gln Ala Leu Ala Asp Lys Tyr Pro Ser 50 55 60 Glu Asn Leu Leu Leu Val Thr His Gly Glu Gly Val Gly Thr Ser Val 65 70 75 80 Ala Thr Phe Leu Lys Gly Ala Val Val Tyr Glu Val Lys Tyr Cys Ala 85 90 95 Tyr Ser Gln Ala Thr Arg Arg Ile Ser Tyr Gly Glu Gly Glu Ser Phe 100 105 110 Thr Ala Gly Thr Phe Gln Leu Val Thr Ala Ser Asp Gln Thr Gly Ile 115 120 125 Gly Tyr Tyr Thr Ser Ser Ser Leu Ser Asp Gly Val 130 135 140 <210> 37 <211> 348 <212> DNA <213> Sueda japonica <220> <221> CDS <222> (1). . (246) <400> 37 atc att gct ccc cta gct att ggt tttg
atc gtt ggt gcc aac atc tta 48 Ile Ile Ala Pro Leu Ala Ile Gly Leu
Ile Val Gly Ala Asn Ile Leu 15
10 15 gcc gga ggt gca ttt gat ggt gcc tca
atg aac cct gcc gtc tct ttt 96 Ala Gly Gly Ala Phe Asp Gly Ala Ser
Met Asn Pro Ala Val Ser Phe 20 25
30 ggc ccc gcc gtg gtt agc tgg agc tgg
gcc aac cac tgg gtc tac tgg 144 Gly Pro Ala Val Val Ser Trp Ser Trp
Ala Asn His Trp Val Tyr Trp 35 40
45 gca ggc cca ctc att ggt ggt gga ctt
gct ggt ctc gtt tat gag ttt 192 Ala Gly Pro Leu Ile Gly Gly Gly Leu
Ala Gly Leu Val Tyr Glu Phe 50 55
60 atc ttt att ggt cac caa gag cca gct
tcc gct gac tac cag aga ctc 240 Ile Phe Ile Gly His Gln Glu Pro Ala
Ser Ala Asp Tyr Gln Arg Leu 65 70
75 80 tct gct taagaatttt aattctttgc cctagggaaa aatgtttcat gcatgtattt 296 Ser Ala tggtattttg ttgggtctaa aattttatga agggaaaaaa aaaaaaaaaa aa 348 <210> 38 <211> 82 <212> PRT <213> Sueada japonica <400> 38 Ile Ile Ala Pro Leu Ala Ile Gly Leu Ile Val Gly Ala Asn Ile Leu 1 5 10 15 Ala Gly Gly Ala Phe Asp Gly Ala Ser Met Asn Pro Ala Val Ser Phe 20 25 30 Gly Pro Ala Val Val Ser Trp Ser Trp Ala Asn His Trp Val Tyr Trp 35 40 45 Ala Gly Pro Leu Ile Gly Gly Gly Leu Ala Gly Leu Val Tyr Glu Phe 50 55 60 Ile Phe Ile Gly His Gln Glu Pro Ala Ser Ala Asp Tyr Gln Arg Leu 65 70 75 80 Ser Ala  <210> 39 <211> 1602 <212> DNA <213> Sueada japonica <220> <221> CDS <222> (1) .. (1419) <400> 39 cac acc gtt gat tta acc att gaa gct atg atg ctc gat tct caa gct 48 His Thr Val Asp Leu Thr Ile Glu Ala Met Met Leu Asp Ser Gln Ala 1 5 10 15 tct gat ctt gac aaa gaa gaa cgt cct gag att ctt tca atg ctt ccg 96 Ser Asp Leu Asp Lys Glu Glu Arg Pro Glu Ile Leu Ser Met Leu Pro 20 25 30 cct ctt gaa gga aaa tgc ctc ttg gaa ctt ggg gct ggt att ggt cgt 144 Pro Leu Glu Gly Lys Cys Leu Leu Glu Leu Gly Ala Gly Ile Gly Arg 35 40 45 ttt act ggt gaa ttg gct gag aaa gct ggc cag gtt att gct ctg gat 192 Phe Thr Gly Glu Leu Ala Glu Lys Ala Gly Gln Val Ile Ala Leu Asp 50 55 60 ttc att gag agt gct atc aag aag aat gaa gta atc aat ggg cac tac 240 Phe Ile Glu Ser Ala Ile Lys Lys Asn Glu Val Ile Asn Gly His Tyr 65 70 75 80 aaa aat gtc aag ttt atg tgt gct gat gtg act tct ccc act ctc agt 288 Lys Asn Val Lys Phe Met Cys Ala Asp Val Thr Ser Pro Thr Leu Ser 85 90 95 ttc cca cca cat tca ttg gat gtg ata ttc tcc aat tgg tta ctc atg 336 Phe Pro Pro His Ser Leu Asp Val Ile Phe Ser Asn Trp Leu Leu Met 100 105 110 tat ctt tct gat gaa gag gtg gaa aat ttg gtt gaa aga atg ttg aaa 384 Tyr Leu Ser Asp Glu Glu Val Glu Asn Leu Val Glu Arg Met Leu Lys 115 120 125 tgg ttg aag cca ggg ggt tac att ttc ttc aga gaa tct tgt ttc cat 432 Trp Leu Lys Pro Gly Gly Tyr Ile Phe Phe Arg Glu Ser Cys Phe His 130 135 140 caa tct ggg gat cac aaa cgc aaa agc aat ccc acc cac tac cgt gaa 480 Gln Ser Gly Asp His Lys Arg Lys Ser Asn Pro Thr His Tyr Arg Glu 145 150 155 160 cct agg ttc tac act aag gcc ttc aaa gag tgt cat ttg caa gat gga 528 Pro Arg Phe Tyr Thr Lys Ala Phe Lys Glu Cys His Leu Gln Asp Gly 165 170 175 tct gga aac tct tat gag ctc tcc cta ctt agc tgc aaa tgt att gga 576 Ser Gly Asn Ser Tyr Glu Leu Ser Leu Leu Ser Cys Lys Cys Ile Gly 180 185 190 gct tat gtc aga aac aag aaa aac cag aac cag att agt tgg ttg tgg 624 Ala Tyr Val Arg Asn Lys Lys Asn Gln Asn Gln Ile Ser Trp Leu Trp 195 200 205 caa aaa gtt gat tct aag gat gat aag ggg ttc cag cga ttt ctg gat 672 Gln Lys Val Asp Ser Lys Asp Asp Lys Gly Phe Gln Arg Phe Leu Asp 210 215 220 act agc cag tac aag tgt aat agc att ctg cga tat gag cgt gta ttt 720 Thr Ser Gln Tyr Lys Cys Asn Ser Ile Leu Arg Tyr Glu Arg Val Phe 225 230 235 240 ggc cct ggt tat gtt agc act gga gga tat gaa acc acc aaa gag ttt 768 Gly Pro Gly Tyr Val Ser Thr Gly Gly Tyr Glu Thr Thr Lys Glu Phe 245 250 255 gtg tca atg ctg gac ttg aag cct ggc cag aag gtc ctg gat gtt ggt 816 Val Ser Met Leu Asp Leu Lys Pro Gly Gln Lys Val Leu Asp Val Gly 260 265 270 tgt gga att ggt gga ggt gac ttt tac atg gcg gag acc ttt gat gtt 864 Cys Gly Ile Gly Gly Gly Asp Phe Tyr Met Ala Glu Thr Phe Asp Val 275 280 285 gag gtt gtt gga ttt gat ctc tcc gtt aat atg att tcc ttt gcc ctt 912 Glu Val Val Gly Phe Asp Leu Ser Val Asn Met Ile Ser Phe Ala Leu 290 295 300 gag cgt tct att ggg ctt aaa tgt gct gtt gag ttt gag gta gca gat 960 Glu Arg Ser Ile Gly Leu Lys Cys Ala Val Glu Phe Glu Val Ala Asp 305 310 315 320 tgc acc aag ata aac tac cct gat aac tct ttt gat gtc atc tat agc 1008 Cys Thr Lys Ile Asn Tyr Pro Asp Asn Ser Phe Asp Val Ile Tyr Ser 325 330 335 cgt gac acc att ctg cat att cag gac aag cct gcg ttg ttt aga tcc 1056 Arg Asp Thr Ile Leu His Ile Gln Asp Lys Pro Ala Leu Phe Arg Ser 340 345 350 ttc tac aaa tgg ttg aag cca gga ggt aaa gtt cta atc agt gat tac 1104 Phe Tyr Lys Trp Leu Lys Pro Gly Gly Lys Val Leu Ile Ser Asp Tyr 355 360 365 tgc aag aaa gct ggt cca ccc tca cct gaa ttc gcc gct tac att aag 1152 Cys Lys Lys Ala Gly Pro Pro Ser Pro Glu Phe Ala Ala Tyr Ile Lys 370 375 380 cag agg gga tat gat ctc cat gat gta aag gaa tat ggg cag atg ctt 1200 Gln Arg Gly Tyr Asp Leu His Asp Val Lys Glu Tyr Gly Gln Met Leu 385 390 395 400 aaa gat gct gga ttt gtt gat gtt ctt gcc gag gat aga act gag cag 1248 Lys Asp Ala Gly Phe Val Asp Val Leu Ala Glu Asp Arg Thr Glu Gln 405 410 415 ttc att cga gtt cta cgg aag gaa cta gag act gtt gag aag gaa aag 1296 Phe Ile Arg Val Leu Arg Lys Glu Leu Glu Thr Val Glu Lys Glu Lys 420 425 430 gat gtg ttc att agt gat ttc tct gag gag gat tac aat gac att gtt 1344 Asp Val Phe Ile Ser Asp Phe Ser Glu Glu Asp Tyr Asn Asp Ile Val 435 440 445 gga ggt tgg aat gat aag ttg cgg agg act gcc aag ggt gag caa cga 1392 Gly Gly Trp Asn Asp Lys Leu Arg Arg Thr Ala Lys Gly Glu Gln Arg 450 455 460 tgg ggt ctg ttc gtt gcc aag aag aag tgaagaatca gttgccgcac 1439 Trp Gly Leu Phe Val Ala Lys Lys Lys 465 470 tggcactgtc gatttcctag tattaatctt caatgttttc atgtaatgta cttctacatg 1499 taaaattgcc aataagttgc atttcgcaga ctgtaagatg attaatcata ttttatcttt 1559 taattaatca tggatttatg caaaaaaaaa aaaaaaaaaa aaa 1602 <210> 40 <211> 473 <212> PRT <213> Sueada japonica <400> 40 His Thr Val Asp Leu Thr Ile Glu Ala Met Met Leu Asp Ser Gln Ala 1 5 10 15 Ser Asp Leu Asp Lys Glu Glu Arg Pro Glu Ile Leu Ser Met Leu Pro 20 25 30 Pro Leu Glu Gly Lys Cys Leu Leu Glu Leu Gly Ala Gly Ile Gly Arg 35 40 45 Phe Thr Gly Glu Leu Ala Glu Lys Ala Gly Gln Val Ile Ala Leu Asp 50 55 60 Phe Ile Glu Ser Ala Ile Lys Lys Asn Glu Val Ile Asn Gly His Tyr 65 70 75 80 Lys Asn Val Lys Phe Met Cys Ala Asp Val Thr Ser Pro Thr Leu Ser 85 90 95 Phe Pro Pro His Ser Leu Asp Val Ile Phe Ser Asn Trp Leu Leu Met 100 105 110 Tyr Leu Ser Asp Glu Glu Val Glu Asn Leu Val Glu Arg Met Leu Lys 115 120 125 Trp Leu Lys Pro Gly Gly Tyr Ile Phe Phe Arg Glu Ser Cys Phe His 130 135 140 Gln Ser Gly Asp His Lys Arg Lys Ser Asn Pro Thr His Tyr Arg Glu 145 150 155 160 Pro Arg Phe Tyr Thr Lys Ala Phe Lys Glu Cys His Leu Gln Asp Gly 165 170 175 Ser Gly Asn Ser Tyr Glu Leu Ser Leu Leu Ser Cys Lys Cys Ile Gly 180 185 190 Ala Tyr Val Arg Asn Lys Lys Asn Gln Asn Gln Ile Ser Trp Leu Trp 195 200 205 Gln Lys Val Asp Ser Lys Asp Asp Lys Gly Phe Gln Arg Phe Leu Asp 210 215 220 Thr Ser Gln Tyr Lys Cys Asn Ser Ile Leu Arg Tyr Glu Arg Val Phe 225 230 235 240 Gly Pro Gly Tyr Val Ser Thr Gly Gly Tyr Glu Thr Thr Lys Glu Phe 245 250 255 Val Ser Met Leu Asp Leu Lys Pro Gly Gln Lys Val Leu Asp Val Gly 260 265 270 Cys Gly Ile Gly Gly Gly Asp Phe Tyr Met Ala Glu Thr Phe Asp Val 275 280 285 Glu Val Val Gly Phe Asp Leu Ser Val Asn Met Ile Ser Phe Ala Leu 290 295 300 Glu Arg Ser Ile Gly Leu Lys Cys Ala Val Glu Phe Glu Val Ala Asp 305 310 315 320 Cys Thr Lys Ile Asn Tyr Pro Asp Asn Ser Phe Asp Val Ile Tyr Ser 325 330 335 Arg Asp Thr Ile Leu His Ile Gln Asp Lys Pro Ala Leu Phe Arg Ser 340 345 350 Phe Tyr Lys Trp Leu Lys Pro Gly Gly Lys Val Leu Ile Ser Asp Tyr 355 360 365 Cys Lys Lys Ala Gly Pro Pro Ser Pro Glu Phe Ala Ala Tyr Ile Lys 370 375 380 Gln Arg Gly Tyr Asp Leu His Asp Val Lys Glu Tyr Gly Gln Met Leu 385 390 395 400 Lys Asp Ala Gly Phe Val Asp Val Leu Ala Glu Asp Arg Thr Glu Gln 405 410 415 Phe Ile Arg Val Leu Arg Lys Glu Leu Glu Thr Val Glu Lys Glu Lys 420 425 430 Asp Val Phe Ile Ser Asp Phe Ser Glu Glu Asp Tyr Asn Asp Ile Val 435 440 445 Gly Gly Trp Asn Asp Lys Leu Arg Arg Thr Ala Lys Gly Glu Gln Arg 450 455 460 Trp Gly Leu Phe Val Ala Lys Lys Lys 465 470 <210> 41 <211> 1251 <212> DNA <213> Salsola komarovii <220> <221> CDS <222> (1) .. (933) <400> 41 cag cca ttt ggc aca att aat gga tca ctt cgt gtt act gta caa ggt 48 Gln Pro Phe Gly Thr Ile Asn Gly Ser Leu Arg Val Thr Val Gln Gly 1 5 10 15 gag gtc att gaa caa tct ttt gga gag gag cac ttg tgt ttt aga aca 96 Glu Val Ile Glu Gln Ser Phe Gly Glu Glu His Leu Cys Phe Arg Thr 20 25 30 tta cag cgg tac aca gct gcc aca ctt gag cat gga atg cat cca cca 144 Leu Gln Arg Tyr Thr Ala Ala Thr Leu Glu His Gly Met His Pro Pro 35 40 45 atc tct cct aaa cca gaa tgg cgt gca ctt ttg gac gag atg gct gtt 192 Ile Ser Pro Lys Pro Glu Trp Arg Ala Leu Leu Asp Glu Met Ala Val 50 55 60 gtt gcc acc aag gaa tac cgc tct gtt gtt ttt cat gag cct cgc ttt 240 Val Ala Thr Lys Glu Tyr Arg Ser Val Val Phe His Glu Pro Arg Phe 65 70 75 80 gtc gag tac ttc cgc agt gct aca cca gag aca gag tat ggg cgt atg 288 Val Glu Tyr Phe Arg Ser Ala Thr Pro Glu Thr Glu Tyr Gly Arg Met 85 90 95 aat att gga agc cgt cct gca aag aga aag cca gga gga gga att gaa 336 Asn Ile Gly Ser Arg Pro Ala Lys Arg Lys Pro Gly Gly Gly Ile Glu 100 105 110 act ctg cgt gca att cct tgg ata ttt tcg tgg aca caa acc agg ttt 384 Thr Leu Arg Ala Ile Pro Trp Ile Phe Ser Trp Thr Gln Thr Arg Phe 115 120 125 cat tta cct gtg tgg ctt ggg gtt gga gca gct ttt aag cat gcc ctt 432 His Leu Pro Val Trp Leu Gly Val Gly Ala Ala Phe Lys His Ala Leu 130 135 140 gac aag gac att aag aat ctt tcg ata ctc aag gcc atg tat aat gag 480 Asp Lys Asp Ile Lys Asn Leu Ser Ile Leu Lys Ala Met Tyr Asn Glu 145 150 155 160 tgg ccg ttc ttc aga gtg act att gat ctc tta gaa atg gtt ttc act 528 Trp Pro Phe Phe Arg Val Thr Ile Asp Leu Leu Glu Met Val Phe Thr 165 170 175 aaa gga gac cct gga att gct gct tta tat gac aag ctt ctg gtg gca 576 Lys Gly Asp Pro Gly Ile Ala Ala Leu Tyr Asp Lys Leu Leu Val Ala 180 185 190 gag gat ttg aag ccc ttt ggg gaa aag ttg agg aaa agt ttc gaa gat 624 Glu Asp Leu Lys Pro Phe Gly Glu Lys Leu Arg Lys Ser Phe Glu Asp 195 200 205 acc aaa ctc ctt ctc ctt aag gtt gct ggg cac aag gag tta ctg gaa 672 Thr Lys Leu Leu Leu Leu Lys Val Ala Gly His Lys Glu Leu Leu Glu 210 215 220 gga gat cct tac ttg aaa cag aga ctc cga ctt cgt gat cct tac att 720 Gly Asp Pro Tyr Leu Lys Gln Arg Leu Arg Leu Arg Asp Pro Tyr Ile 225 230 235 240 aca acc ctt aat gtt ttc caa gca tat act ctg aag cgg atc cgt gat 768 Thr Thr Leu Asn Val Phe Gln Ala Tyr Thr Leu Lys Arg Ile Arg Asp 245 250 255 ccc aat ttc cat gta gct gaa ggg cca cac tta tcc aag gaa gta ttg 816 Pro Asn Phe His Val Ala Glu Gly Pro His Leu Ser Lys Glu Val Leu 260 265 270 gaa tca aac aat gct gag ctt gtg aag ctc aat cct act agt gag tat 864 Glu Ser Asn Asn Ala Glu Leu Val Lys Leu Asn Pro Thr Ser Glu Tyr 275 280 285 cct cct ggc ctt gag gac acc ctt atc ttg acc atg aag ggt att gct 912 Pro Pro Gly Leu Glu Asp Thr Leu Ile Leu Thr Met Lys Gly Ile Ala 290 295 300 gct ggc atg cag aac acc ggt taactgacac gtgttgcacg tctattgcaa 963 Ala Gly Met Gln Asn Thr Gly 305 310 ctattcctca actccttctg gtttggggat ccgggctcgg agatagccat cgttggtgat 1023 gtgctgtatg agcacctaat tgtattcaaa gtctgtattt caagtctatt gtatttgtat 1083 tttgttcttc tgtatgtttt tgttatttct acttatggtt gggttgtgtc acttgtgact 1143 aatacccgac tgtgtaataa atggttgttg tactgatgaa cagtttgttt tcttctacgt 1203 gagttatatt gatgagttta tcttttatta aaaaaaaaaa aaaaaaaa 1251 <210> 42 <211> 311 <212> PRT <213> Salsola komarovii <400> 42 Gln Pro Phe Gly Thr Ile Asn Gly Ser Leu Arg Val Thr Val Gln Gly 1 5 10 15 Glu Val Ile Glu Gln Ser Phe Gly Glu Glu His Leu Cys Phe Arg Thr 20 25 30 Leu Gln Arg Tyr Thr Ala Ala Thr Leu Glu His Gly Met His Pro Pro 35 40 45 Ile Ser Pro Lys Pro Glu Trp Arg Ala Leu Leu Asp Glu Met Ala Val 50 55 60 Val Ala Thr Lys Glu Tyr Arg Ser Val Val Phe His Glu Pro Arg Phe 65 70 75 80 Val Glu Tyr Phe Arg Ser Ala Thr Pro Glu Thr Glu Tyr Gly Arg Met 85 90 95 Asn Ile Gly Ser Arg Pro Ala Lys Arg Lys Pro Gly Gly Gly Ile Glu 100 105 110 Thr Leu Arg Ala Ile Pro Trp Ile Phe Ser Trp Thr Gln Thr Arg Phe 115 120 125 His Leu Pro Val Trp Leu Gly Val Gly Ala Ala Phe Lys His Ala Leu 130 135 140 Asp Lys Asp Ile Lys Asn Leu Ser Ile Leu Lys Ala Met Tyr Asn Glu 145 150 155 160 Trp Pro Phe Phe Arg Val Thr Ile Asp Leu Leu Glu Met Val Phe Thr 165 170 175 Lys Gly Asp Pro Gly Ile Ala Ala Leu Tyr Asp Lys Leu Leu Val Ala 180 185 190 Glu Asp Leu Lys Pro Phe Gly Glu Lys Leu Arg Lys Ser Phe Glu Asp 195 200 205 Thr Lys Leu Leu Leu Leu Lys Val Ala Gly His Lys Glu Leu Leu Glu 210 215 220 Gly Asp Pro Tyr Leu Lys Gln Arg Leu Arg Leu Arg Asp Pro Tyr Ile 225 230 235 240 Thr Thr Leu Asn Val Phe Gln Ala Tyr Thr Leu Lys Arg Ile Arg Asp 245 250 255 Pro Asn Phe His Val Ala Glu Gly Pro His Leu Ser Lys Glu Val Leu 260 265 270 Glu Ser Asn Asn Ala Glu Leu Val Lys Leu Asn Pro Thr Ser Glu Tyr 275 280 285 Pro Pro Gly Leu Glu Asp Thr Leu Ile Leu Thr Met Lys Gly Ile Ala 290 295 300 Ala Gly Met Gln Asn Thr Gly 305 310 <210> 43 <211> 637 <212> DNA <213> Avicennia marina <220> <221> CDS <222> (1) .. (339) <400> 43 caa tac ttg gta aat gaa gtg aag aaa act gtt cag ggg cgt gct caa 48 Gln Tyr Leu Val Asn Glu Val Lys Lys Thr Val Gln Gly Arg Ala Gln 1 5 10 15 ctt ggt gtg gaa gca ttt gct gat gcg ctt ctt gtg gtt cca aag acg 96 Leu Gly Val Glu Ala Phe Ala Asp Ala Leu Leu Val Val Pro Lys Thr 20 25 30 ctt gcc gag aac tct ggc ctt gat acc cag gat ttg att att gaa ctt 144 Leu Ala Glu Asn Ser Gly Leu Asp Thr Gln Asp Leu Ile Ile Glu Leu 35 40 45 acg gga gaa tat gaa aaa ggg aat gtg gta gga ctt aat cta cac aca 192 Thr Gly Glu Tyr Glu Lys Gly Asn Val Val Gly Leu Asn Leu His Thr 50 55 60 gga gaa cct ata gat cct caa atg gag ggt atc ttt gac aat tat tcc 240 Gly Glu Pro Ile Asp Pro Gln Met Glu Gly Ile Phe Asp Asn Tyr Ser 65 70 75 80 gtg aag cgt cag atc ata aac tca ggc ccc gtt att gca tct cag ctg 288 Val Lys Arg Gln Ile Ile Asn Ser Gly Pro Val Ile Ala Ser Gln Leu 85 90 95 cta ctt gtc gac gag gtt att cgt gct ggt cgt aac atg cgt aaa ccg 336 Leu Leu Val Asp Glu Val Ile Arg Ala Gly Arg Asn Met Arg Lys Pro 100 105 110 aat tagctttcac cctagttttt gtgatgttgg tgaagatggt aattttattt 389 Asn aggtagggtc atggttcctt ttgtttagcc taagcactat gtattcattg ccacttgaga 449 tttgaatttt gatcatcagg cggttgaact tttcgcctgt tacaaattgc accagaaatt 509 attcgaccat gggtatgcat ctacttgtgt tgtacctgac ttggctaagt tatttgaaga 569 tacactctgt gctcagcaaa gaattggaaa aaaaggaatt gatttcatca aaaaaaaaaa 629 aaaaaaaa 637 <210> 44 <211> 113 <212> PRT <213> Avicennia marina <400> 44 Gln Tyr Leu Val Asn Glu Val Lys Lys Thr Val Gln Gly Arg Ala Gln 1 5 10 15 Leu Gly Val Glu Ala Phe Ala Asp Ala Leu Leu Val Val Pro Lys Thr 20 25 30 Leu Ala Glu Asn Ser Gly Leu Asp Thr Gln Asp Leu Ile Ile Glu Leu 35 40 45 Thr Gly Glu Tyr Glu Lys Gly Asn Val Val Gly Leu Asn Leu His Thr 50 55 60 Gly Glu Pro Ile Asp Pro Gln Met Glu Gly Ile Phe Asp Asn Tyr Ser 65 70 75 80 Val Lys Arg Gln Ile Ile Asn Ser Gly Pro Val Ile Ala Ser Gln Leu 85 90 95 Leu Leu Val Asp Glu Val Ile Arg Ala Gly Arg Asn Met Arg Lys Pro 100 105 110 Asn  <210> 45 <211> 741 <212> DNA <213> Avicennia marina <220> <221> CDS <222> (3) .. (293) <400> 45 aa gag atc aat tgt ctt gaa tgg gag aac ttt gct ttc cat ccc agc 47 Glu Ile Asn Cys Leu Glu Trp Glu Asn Phe Ala Phe His Pro Ser 1 5 10 15 cca ctc att gtt ctt gtt ttt gaa aga tac aac agg gca agt gat aac 95 Pro Leu Ile Val Leu Val Phe Glu Arg Tyr Asn Arg Ala Ser Asp Asn 20 25 30 tgg aaa gct ttg aag gag ttg gaa aag gcg gca gaa gtt tac tgg aag 143 Trp Lys Ala Leu Lys Glu Leu Glu Lys Ala Ala Glu Val Tyr Trp Lys 35 40 45 gca aaa gat cga ctg cct cct cgg acg gtc aag ata gat ata aac atc 191 Ala Lys Asp Arg Leu Pro Pro Arg Thr Val Lys Ile Asp Ile Asn Ile 50 55 60 gaa agg gat tta gca tat gca ctc aag gtt aaa gaa tgc ccg cag ata 239 Glu Arg Asp Leu Ala Tyr Ala Leu Lys Val Lys Glu Cys Pro Gln Ile 65 70 75 ctg ttc tta cgc gga aac agg ata tta tac aga gag aaa ggt agc cca 287 Leu Phe Leu Arg Gly Asn Arg Ile Leu Tyr Arg Glu Lys Gly Ser Pro 80 85 90 95 ttt ctc tgatattgca tgtacatcag atctttcaat ctgcaccaga accaattgag 343 Phe Leu tttaccatca tttccagaaa ttagatcatc ggatgaattg gttcagatga tcgcgcattt 403 ctattacaat gcaaaaaagc cttcgtgcat cgatgatgca gctttctctt caccacatca 463 ctgaaggtga ggttgtcaaa tggaatccag catcagtcat tagggaggac tgaagctgta 523 cggagggaag tggtttaaat tcagattgga tctttgaagt gggcagtggt gattgaaacg 583 ccaaaagttt ctgaggaata accttgttgg gattttgcag tgaactgtag taactttctc 643 gcatgtaaaa ctagactttc atcaatcaac caccaaccct tttatgtata tgaaacctat 703 gaggttgaaa tttctagtta aaaaaaaaaa aaaaaaaa 741 <210> 46 <211> 97 <212> PRT <213> Avicennia marina <400> 46 Glu Ile Asn Cys Leu Glu Trp Glu Asn Phe Ala Phe His Pro Ser Pro 1 5 10 15 Leu Ile Val Leu Val Phe Glu Arg Tyr Asn Arg Ala Ser Asp Asn Trp 20 25 30 Lys Ala Leu Lys Glu Leu Glu Lys Ala Ala Glu Val Tyr Trp Lys Ala 35 40 45 Lys Asp Arg Leu Pro Pro Arg Thr Val Lys Ile Asp Ile Asn Ile Glu 50 55 60 Arg Asp Leu Ala Tyr Ala Leu Lys Val Lys Glu Cys Pro Gln Ile Leu 65 70 75 80 Phe Leu Arg Gly Asn Arg Ile Leu Tyr Arg Glu Lys Gly Ser Pro Phe 85 90 95 Leu  <210> 47 <211> 983 <212> DNA <213> Salsola komarovii <220> <221> CDS <222> (1) .. (762) <400> 47 atg ttc ctt cat cac cac ttt tca tct tca tct tct tct ttt ctt ctt 48 Met Phe Leu His His His Phe Ser Ser Ser Ser Ser Ser Phe Leu Leu 1 5 10 15 ctc ttc ttc tct ctc cta ata ttc ctt tca tct gct aat ctt tat cat 96 Leu Phe Phe Ser Leu Leu Ile Phe Leu Ser Ser Ala Asn Leu Tyr His 20 25 30 cag aat caa gga tct tgt agt gac ttt gaa tca gaa cca tca atg gct 144 Gln Asn Gln Gly Ser Cys Ser Asp Phe Glu Ser Glu Pro Ser Met Ala 35 40 45 act ctt ggt gga ttg cgc gaa tcc cat ggt gct tct aat gat gct gag 192 Thr Leu Gly Gly Leu Arg Glu Ser His Gly Ala Ser Asn Asp Ala Glu 50 55 60 att gaa acc ctt gct cgc ttt gct gtt gat gaa cac aac aaa aaa gag 240 Ile Glu Thr Leu Ala Arg Phe Ala Val Asp Glu His Asn Lys Lys Glu 65 70 75 80 aat gca ttg ttg gag ttt gca agg gtt gta aag gca aag gaa cag gtg 288 Asn Ala Leu Leu Glu Phe Ala Arg Val Val Lys Ala Lys Glu Gln Val 85 90 95 gtt gcg ggt aca ttg cat cac ttc act atc gaa gca att gaa gcg ggc 336 Val Ala Gly Thr Leu His His Phe Thr Ile Glu Ala Ile Glu Ala Gly 100 105 110 aag aag aag ctc tac gaa gcg aag gtg
tgg gtg aag cca tgg atg aac 384 Lys Lys Lys Leu Tyr Glu Ala Lys Val
Trp Val Lys Pro Trp Met Asn 115 120
125 ttt aag gag ctg cag gaa ttt aag cat
gct gat gaa tcc cct tca atc 432 Phe Lys Glu Leu Gln Glu Phe Lys His
Ala Asp Glu Ser Pro Ser Ile 130 135
140 act cct tcc gac ctc ggc gct aat aga
ga aggg cat tct gga gga tgg 480 Thr Pro Ser Asp Leu Gly Ala Asn Arg
Glu Gly His Ser Gly Gly Trp 145 150
155 160 aaa gat gtg cct gtc cat gac cct gaa
gtg caa aat gca gca aat cat 528 Lys Asp Val Pro Val His His Asp Pro Glu
Val Gln Asn Ala Ala Asn His 165
170 175 gct ctt aag acc ttg caa caa aga tcc
aac tcc tta ttt cct tat ga 576 Ala Leu Lys Thr Leu Gln Gln Arg Ser
Asn Ser Leu Phe Pro Tyr Glu 180 185
190 ctg cag gaa gtt gct cat gct agg gct
gag gtt ctg gaa gac act gcg 624 Leu Gln Glu Val Ala His Ala Arg Ala
Glu Val Leu Glu Asp Thr Ala 195 200
205 aag ttt aac ctg cac ctc aag gtg aag
aga gga aac aag gat gag ttt 672 Lys Phe Asn Leu His Leu Lys Val Lys
Arg Gly Asn Lys Asp Glu Phe 210 215
220 ttc aat gtg gag gtg cac aaa aac agc gaa gga aac tac aac ctt aat 720 Phe Asn Val Glu Val His Lys Asn Ser Glu Gly Asn Tyr Asn Leu Asn 225 230 235 240 cag atg ggg aac gtt gag ccc gag gtt gag aaa agt agt gtt 762 Gln Met Gly Asn Val Glu Pro Glu Val Glu Lys Ser Ser Val 245 250 tagactcgtt gagggtgttg taagtactcg ttcgtaactt ttctgatggt caggcaagta 822 tggagtaagg actagactac tagtactagt aagtacagct gacttggttt gagtaaaata 882 acctcgactt tggttgcacc atcatatctt gtatgtttat ggctttgtca atgtattgta 942 agtgaagatt gtttgcttga tctaaaaaaa aaaaaaaaaa a 983 <210> 48 <211> 254 <212> PRT <213> Salsola komarovii <400> 48 Met Phe Leu His His His Phe Ser Ser Ser Ser Ser Ser Phe Leu Leu 1 5 10 15 Leu Phe Phe Ser Leu Leu Ile Phe Leu Ser Ser Ala Asn Leu Tyr His 20 25 30 Gln Asn Gln Gly Ser Cys Ser Asp Phe Glu Ser Glu Pro Ser Met Ala 35 40 45 Thr Leu Gly Gly Leu Arg Glu Ser His Gly Ala Ser Asn Asp Ala Glu 50 55 60 Ile Glu Thr Leu Ala Arg Phe Ala Val Asp Glu His Asn Lys Lys Glu 65 70 75 80 Asn Ala Leu Leu Glu Phe Ala Arg Val Val Lys Ala Lys Glu Gln Val 85 90 95 Val Ala Gly Thr Leu His His Phe Thr Ile Glu Ala Ile Glu Ala Gly 100 105 110 Lys Lys Lys Leu Tyr Glu Ala Lys Val Trp Val Lys Pro Trp Met Asn 115 120 125 Phe Lys Glu Leu Gln Glu Phe Lys His Ala Asp Glu Ser Pro Ser Ile 130 135 140 Thr Pro Ser Asp Leu Gly Ala Asn Arg Glu Gly His Ser Gly Gly Trp 145 150 155 160 Lys Asp Val Pro Val His Asp Pro Glu Val Gln Asn Ala Ala Asn His 165 170 175 Ala Leu Lys Thr Leu Gln Gln Arg Ser Asn Ser Leu Phe Pro Tyr Glu 180 185 190 Leu Gln Glu Val Ala His Ala Arg Ala Glu Val Leu Glu Asp Thr Ala 195 200 205 Lys Phe Asn Leu His Leu Lys Val Lys Arg Gly Asn Lys Asp Glu Phe 210 215 220 Phe Asn Val Glu Val His Lys Asn Ser Glu Gly Asn Tyr Asn Leu Asn 225 230 235 240 Gln Met Gly Asn Val Glu Pro Glu Val Glu Lys Ser Ser Val 245 250 <210> 49 <211> 543 <212> DNA <213> Salsola komarovii <220> <221> CDS <222> (3) .. (389) <400> 49 aa aat aag gtt gac tta gct cga gat ttc acc ttc ata gac gac gtc 47 Asn Lys Val Asp Leu Ala Arg Asp Phe Thr Phe Ile Asp Asp Val 1 5 10 15 gta aag ggg tgc tta ggt tca ctg gat tct tcc ggt aag agt acc ggt 95 Val Lys Gly Cys Leu Gly Ser Leu Asp Ser Ser Gly Lys Ser Thr Gly 20 25 30 agc ggc ggt aaa aaa cgt ggg ccc gct ccg tac aga atc tac aac ttg 143 Ser Gly Gly Lys Lys Arg Gly Pro Ala Pro Tyr Arg Ile Tyr Asn Leu 35 40 45 ggg aac act caa ccg gtc act gta ccg aca ctt gtc ggt atc cta gag 191 Gly Asn Thr Gln Pro Val Thr Val Pro Thr Leu Val Gly Ile Leu Glu 50 55 60 aag cat ctc aaa gtt aag gcc aag aag aat gtg gtt gag atg ccc gga 239 Lys His Leu Lys Val Lys Ala Lys Lys Asn Val Val Glu Met Pro Gly 65 70 75 aat ggt gac gtg ccc ttc aca cat gcg aat atc tct ttg gcc cga aaa 287 Asn Gly Asp Val Pro Phe Thr His Ala Asn Ile Ser Leu Ala Arg Lys 80 85 90 95 gat ttc ggg tat aaa ccc act acc gat ttg caa acc ggg ttg aaa aag 335 Asp Phe Gly Tyr Lys Pro Thr Thr Asp Leu Gln Thr Gly Leu Lys Lys 100 105 110 ttt gtt aga tgg tat ctc act tat tac ggc tac aac aac ggc aag cct 383 Phe Val Arg Trp Tyr Leu Thr Tyr Tyr Gly Tyr Asn Asn Gly Lys Pro 115 120 125 gta aat taatatataa atataagtaa tatttttttt ctcttttttt ataaattaca 439 Val Asn gaattatttt ttttgggtgg tttatgaatt ttgttggata atatggggat tctttttttc 499 taaatgggaa aaataagaat ccaaggaaaa aaaaaaaaaa aaaa 543 <210> 50 <211> 129 <212> PRT <213> Salsola komarovii <400> 50 Asn Lys Val Asp Leu Ala Arg Asp Phe Thr Phe Ile Asp Asp Val Val 1 5 10 15 Lys Gly Cys Leu Gly Ser Leu Asp Ser Ser Gly Lys Ser Thr Gly Ser 20 25 30 Gly Gly Lys Lys Arg Gly Pro Ala Pro Tyr Arg Ile Tyr Asn Leu Gly 35 40 45 Asn Thr Gln Pro Val Thr Val Pro Thr Leu Val Gly Ile Leu Glu Lys 50 55 60 His Leu Lys Val Lys Ala Lys Lys Asn Val Val Glu Met Pro Gly Asn 65 70 75 80 Gly Asp Val Pro Phe Thr His Ala Asn Ile Ser Leu Ala Arg Lys Asp 85 90 95 Phe Gly Tyr Lys Pro Thr Thr Asp Leu Gln Thr Gly Leu Lys Lys Phe 100 105 110 Val Arg Trp Tyr Leu Thr Tyr Tyr Gly Tyr Asn Asn Gly Lys Pro Val 115 120 125 Asn  <210> 51 <211> 1219 <212> DNA <213> Sueada japonica <220> <221> CDS <222> (2) .. (871) <400> 51 c aca gga gca aac aaa gga ata gga ctt gaa cta tgc aaa caa cta gct 49 Thr Gly Ala Asn Lys Gly Ile Gly Leu Glu Leu Cys Lys Gln Leu Ala 1 5 10 15 gct aaa gga gtt gta gta gtt ctc act tct aga gat gga aaa aga ggc 97 Ala Lys Gly Val Val Val Val Leu Thr Ser Arg Asp Gly Lys Arg Gly 20 25 30 tta caa gct cat gaa aat ctc att aaa tct gga att aat cct gaa aat 145 Leu Gln Ala His Glu Asn Leu Ile Lys Ser Gly Ile Asn Pro Glu Asn 35 40 45 ctt cac ttt cat cag ctc gat gtt act gac atc act agt att gct gct 193 Leu His Phe His Gln Leu Asp Val Thr Asp Ile Thr Ser Ile Ala Ala 50 55 60 att gct ggt ttc atc aat tcc aaa ttc ggc aaa ctt gat atc ctg gtg 241 Ile Ala Gly Phe Ile Asn Ser Lys Phe Gly Lys Leu Asp Ile Leu Val 65 70 75 80 aac aat gct gga att att gga gat atg gtt aac ttt gat gct tta ata 289 Asn Asn Ala Gly Ile Ile Gly Asp Met Val Asn Phe Asp Ala Leu Ile 85 90 95 gca gca gga ttt ggc act cca aga gaa cag atc aat ctt gag gac agt 337 Ala Ala Gly Phe Gly Thr Pro Arg Glu Gln Ile Asn Leu Glu Asp Ser 100 105 110 ccc ggg aca gta aca cag aca tat gag ctt acg aaa gaa tgc tta caa 385 Pro Gly Thr Val Thr Gln Thr Tyr Glu Leu Thr Lys Glu Cys Leu Gln 115 120 125 aca aat tat tat gga gcg aaa aga acc gtt gaa gct ttg ctt ccg ctt 433 Thr Asn Tyr Tyr Gly Ala Lys Arg Thr Val Glu Ala Leu Leu Pro Leu 130 135 140 ctc aag tta tcc gat tct cca agg att gtc aat gtc tcc tct ttt cta 481 Leu Lys Leu Ser Asp Ser Pro Arg Ile Val Asn Val Ser Ser Phe Leu 145 150 155 160 gga agg ttg acg tat ata cca aat gag acg atc aga ggg gtc cta aga 529 Gly Arg Leu Thr Tyr Ile Pro Asn Glu Thr Ile Arg Gly Val Leu Arg 165 170 175 gat gcc gag agc ctt aca gaa gaa cga ata gat gag att ctg aat gac 577 Asp Ala Glu Ser Leu Thr Glu Glu Arg Ile Asp Glu Ile Leu Asn Asp 180 185 190 atg ctg agg gac ttc aaa gac tgt tca ttc aaa gag aag gga tgg cct 625 Met Leu Arg Asp Phe Lys Asp Cys Ser Phe Lys Glu Lys Gly Trp Pro 195 200 205 aaa aat ctg gca gcc tat ata gtt tca aag gcg gcc ttg agt gca tac 673 Lys Asn Leu Ala Ala Tyr Ile Val Ser Lys Ala Ala Leu Ser Ala Tyr 210 215 220 aca aga ata ctg gct aag aaa tac cca tca atc atg atc aac tgt att 721 Thr Arg Ile Leu Ala Lys Lys Tyr Pro Ser Ile Met Ile Asn Cys Ile 225 230 235 240 tgc cct ggc ttt gtc aaa act gac atc aat gga aac aca gga cac ttg 769 Cys Pro Gly Phe Val Lys Thr Asp Ile Asn Gly Asn Thr Gly His Leu 245 250 255 ccg gtt gaa gaa ggt gca gcg agt ctg gca agg tta gcg ttg atg ccc 817 Pro Val Glu Glu Gly Ala Ala Ser Leu Ala Arg Leu Ala Leu Met Pro 260 265 270 caa att tta cct tct gga cta ttc ttt cag aga act gaa gtt tct tcg 865 Gln Ile Leu Pro Ser Gly Leu Phe Phe Gln Arg Thr Glu Val Ser Ser 275 280 285 ttt gaa taaaacaatt tgcctattca aaccaacacc acatatctat gaagtttcca 921 Phe Glu 290 tttgtaggca tctttacgaa aaaaataaga catctgcaat actgttactg gaaaatgcaa 981 tgtacttttt tcatgtatgc atggcgcagt tatttattct gactgcaaca ataagattct 1041 gttctttcaa ggcactctaa ggaatgctga tgtaccgttc tcaaacaagc agacaagtag 1101 acacgtttga ttgtcatgtc ttcattcgta caatcatttt gtgtttgtat gttgagcatg 1161 tttaactaat tacaagagtg taattaagat caacttttat aaaaaaaaaa aaaaaaaa 1219 <210> 52 <211> 290 <212> PRT <213> Sueada japonica <400> 52 Thr Gly Ala Asn Lys Gly Ile Gly Leu Glu Leu Cys Lys Gln Leu Ala 1 5 10 15 Ala Lys Gly Val Val Val Val Leu Thr Ser Arg Asp Gly Lys Arg Gly 20 25 30 Leu Gln Ala His Glu Asn Leu Ile Lys Ser Gly Ile Asn Pro Glu Asn 35 40 45 Leu His Phe His Gln Leu Asp Val Thr Asp Ile Thr Ser Ile Ala Ala 50 55 60 Ile Ala Gly Phe Ile Asn Ser Lys Phe Gly Lys Leu Asp Ile Leu Val 65 70 75 80 Asn Asn Ala Gly Ile Ile Gly Asp Met Val Asn Phe Asp Ala Leu Ile 85 90 95 Ala Ala Gly Phe Gly Thr Pro Arg Glu Gln Ile Asn Leu Glu Asp Ser 100 105 110 Pro Gly Thr Val Thr Gln Thr Tyr Glu Leu Thr Lys Glu Cys Leu Gln 115 120 125 Thr Asn Tyr Tyr Gly Ala Lys Arg Thr Val Glu Ala Leu Leu Pro Leu 130 135 140 Leu Lys Leu Ser Asp Ser Pro Arg Ile Val Asn Val Ser Ser Phe Leu 145 150 155 160 Gly Arg Leu Thr Tyr Ile Pro Asn Glu Thr Ile Arg Gly Val Leu Arg 165 170 175 Asp Ala Glu Ser Leu Thr Glu Glu Arg Ile Asp Glu Ile Leu Asn Asp 180 185 190 Met Leu Arg Asp Phe Lys Asp Cys Ser Phe Lys Glu Lys Gly Trp Pro 195 200 205 Lys Asn Leu Ala Ala Tyr Ile Val Ser Lys Ala Ala Leu Ser Ala Tyr 210 215 220 Thr Arg Ile Leu Ala Lys Lys Tyr Pro Ser Ile Met Ile Asn Cys Ile 225 230 235 240 Cys Pro Gly Phe Val Lys Thr Asp Ile Asn Gly Asn Thr Gly His Leu 245 250 255 Pro Val Glu Glu Gly Ala Ala Ser Leu Ala Arg Leu Ala Leu Met Pro 260 265 270 Gln Ile Leu Pro Ser Gly Leu Phe Phe Gln Arg Thr Glu Val Ser Ser 275 280 285 Phe Glu 290 <210> 53 <211> 1148 <212> DNA <213> Sueada japonica <220> <221> CDS <222> (3) .. (848) <400> 53 ga agc agg ccg gat atc cat gtt gaa caa gct cat tca gat gat att 47 Ser Arg Pro Asp Ile His Val Glu Gln Ala His Ser Asp Asp Ile 1 5 10 15 act ggg ttg aaa ttc tca tgt gat ggt cgt cat ctg ttg tct aga agt 95 Thr Gly Leu Lys Phe Ser Cys Asp Gly Arg His Leu Leu Ser Arg Ser 20 25 30 ttt gat tgc aca ctt aag gtt tgg gac ttg cgc caa atg aag cgg tct 143 Phe Asp Cys Thr Leu Lys Val Trp Asp Leu Arg Gln Met Lys Arg Ser 35 40 45 ctt aag gtg ttt gat gaa tta cca aat cac tat gct caa acg aat gtc 191 Leu Lys Val Phe Asp Glu Leu Pro Asn His Tyr Ala Gln Thr Asn Val 50 55 60 tca ttt agt cca gat gag cag ctc atc ttg act ggt aca tct gta gaa 239 Ser Phe Ser Pro Asp Glu Gln Leu Ile Leu Thr Gly Thr Ser Val Glu 65 70 75 agg gat agc cca act gga gga ttg ttg tgc ttt tat gat cgg gaa aaa 287 Arg Asp Ser Pro Thr Gly Gly Leu Leu Cys Phe Tyr Asp Arg Glu Lys 80 85 90 95 ctt gaa cta gta tca aaa gtt ggc att tct cct act tgc agt gtt gtg 335 Leu Glu Leu Val Ser Lys Val Gly Ile Ser Pro Thr Cys Ser Val Val 100 105 110 caa tgt gcc tgg cac cca agg ctg aat cag gtt ttt gcc act gct gga 383 Gln Cys Ala Trp His Pro Arg Leu Asn Gln Val Phe Ala Thr Ala Gly 115 120 125 aat aaa agc caa gga ggt aca cat gta ctc tat gat cca acc atg agt 431 Asn Lys Ser Gln Gly Gly Thr His Val Leu Tyr Asp Pro Thr Met Ser 130 135 140 gag aga ggt gct ctt gtg tgt gtt gct cgt gca cca agg atg aaa tca 479 Glu Arg Gly Ala Leu Val Cys Val Ala Arg Ala Pro Arg Met Lys Ser 145 150 155 gtg gat gat ttt gag gtg cag ccg gtt ata cat aac cct cac gca ctt 527 Val Asp Asp Phe Glu Val Gln Pro Val Ile His Asn Pro His Ala Leu 160 165 170 175 ccc ttg ttc aga gat cag cca agc cgc aaa cgt caa aga gag aag att 575 Pro Leu Phe Arg Asp Gln Pro Ser Arg Lys Arg Gln Arg Glu Lys Ile 180 185 190 ctg aag gac cca ata aaa tcc cac aaa cca gag ctt cct atg tca gga 623 Leu Lys Asp Pro Ile Lys Ser His Lys Pro Glu Leu Pro Met Ser Gly 195 200 205 cct ggc cat ggt ggc aga act ggt aca tca tcg ggt agt ttg tta aca 671 Pro Gly His Gly Gly Arg Thr Gly Thr Ser Ser Gly Ser Leu Leu Thr 210 215 220 caa tat ctc ctc aag caa ggg ggc atg ttg aaa gag aca tgg atg gat 719 Gln Tyr Leu Leu Lys Gln Gly Gly Met Leu Lys Glu Thr Trp Met Asp 225 230 235 gaa gat ccc aga gaa gct att ctc aag tat gct gat gct gca gaa aag 767 Glu Asp Pro Arg Glu Ala Ile Leu Lys Tyr Ala Asp Ala Ala Glu Lys 240 245 250 255 gat cca aag ttt att gcc ccg gct tat gct gag act cag ccc aag cca 815 Asp Pro Lys Phe Ile Ala Pro Ala Tyr Ala Glu Thr Gln Pro Lys Pro 260 265 270 gtc ttt gag gat tct gat aag gaa gat gaa gaa taattcatct tttgcagtgg 868 Val Phe Glu Asp Ser Asp Lys Glu Asp Glu Glu 275 280 ttggattaat ttaatttgag aatattatac tgtgtatatt aatagccaat ttttcaggcg 928 aatgatatgc ttctcacatt acatgctgag ttttatttgc tgctacagat tgtagatgaa 988 taggttaatg taaacacaag catagagatt agaatataga aatgattctg tatccaaaac 1048 acaattttat caccagatgg tatcaaaagc tgtattgact gttgagtaat gtcattaacc 1108 actttcactc cccaaaaaaa aaaaaaaaaa aaaaaaaaaa 1148 <210> 54 <211> 282 <212> PRT <213> Sueada japonica <400> 54 Ser Arg Pro Asp Ile His Val Glu Gln Ala His Ser Asp Asp Ile Thr 1 5 10 15 Gly Leu Lys Phe Ser Cys Asp Gly Arg His Leu Leu Ser Arg Ser Phe 20 25 30 Asp Cys Thr Leu Lys Val Trp Asp Leu Arg Gln Met Lys Arg Ser Leu 35 40 45 Lys Val Phe Asp Glu Leu Pro Asn His Tyr Ala Gln Thr Asn Val Ser 50 55 60 Phe Ser Pro Asp Glu Gln Leu Ile Leu Thr Gly Thr Ser Val Glu Arg 65 70 75 80 Asp Ser Pro Thr Gly Gly Leu Leu Cys Phe Tyr Asp Arg Glu Lys Leu 85 90 95 Glu Leu Val Ser Lys Val Gly Ile Ser Pro Thr Cys Ser Val Val Gln 100 105 110 Cys Ala Trp His Pro Arg Leu Asn Gln Val Phe Ala Thr Ala Gly Asn 115 120 125 Lys Ser Gln Gly Gly Thr His Val Leu Tyr Asp Pro Thr Met Ser Glu 130 135 140 Arg Gly Ala Leu Val Cys Val Ala Arg Ala Pro Arg Met Lys Ser Val 145 150 155 160 Asp Asp Phe Glu Val Gln Pro Val Ile His Asn Pro His Ala Leu Pro 165 170 175 Leu Phe Arg Asp Gln Pro Ser Arg Lys Arg Gln Arg Glu Lys Ile Leu 180 185 190 Lys Asp Pro Ile Lys Ser His Lys Pro Glu Leu Pro Met Ser Gly Pro 195 200 205 Gly His Gly Gly Arg Thr Gly Thr Ser Ser Gly Ser Leu Leu Thr Gln 210 215 220 Tyr Leu Leu Lys Gln Gly Gly Met Leu Lys Glu Thr Trp Met Asp Glu 225 230 235 240 Asp Pro Arg Glu Ala Ile Leu Lys Tyr Ala Asp Ala Ala Glu Lys Asp 245 250 255 Pro Lys Phe Ile Ala Pro Ala Tyr Ala Glu Thr Gln Pro Lys Pro Val 260 265 270 Phe Glu Asp Ser Asp Lys Glu Asp Glu Glu 275 280 <210> 55 <211> 1193 <212> DNA <213> Avicennia marina <220> <221> CDS <222> (3) .. (815) <400> 55 gt gca cct gag tta ctt ctt gga gca aag cat tat aca agt gct gtt 47 Ala Pro Glu Leu Leu Leu Gly Ala Lys His Tyr Thr Ser Ala Val 1 5 10 15 gac atg tgg gct gtg ggc tgc att ttt gct gag ctt ctg act cta aag 95 Asp Met Trp Ala Val Gly Cys Ile Phe Ala Glu Leu Leu Thr Leu Lys 20 25 30 cca cta ttt caa ggg caa gaa gta aaa ggg act tct aat cca ttt cag 143 Pro Leu Phe Gln Gly Gln Glu Val Lys Gly Thr Ser Asn Pro Phe Gln 35 40 45 ctt gat caa ctt gac aaa atc ttt aag gtc cta ggt cat ccc acg caa 191 Leu Asp Gln Leu Asp Lys Ile Phe Lys Val Leu Gly His Pro Thr Gln 50 55 60 gaa aag tgg ccc aca cta gcg aat ctt cca cat tgg cag tct gat gtg 239 Glu Lys Trp Pro Thr Leu Ala Asn Leu Pro His Trp Gln Ser Asp Val 65 70 75 caa cgt atc caa ggg ctc aaa tac gac aat act gga ctt tac aat gtt 287 Gln Arg Ile Gln Gly Leu Lys Tyr Asp Asn Thr Gly Leu Tyr Asn Val 80 85 90 95 gtt cat ctc tcc ccc aaa aat cca gca tat gac ctt ctc tca aag atg 335 Val His Leu Ser Pro Lys Asn Pro Ala Tyr Asp Leu Leu Ser Lys Met 100 105 110 ctt gag tat gat cct aga aaa aga ata aca gct aca caa gct ctt gag 383 Leu Glu Tyr Asp Pro Arg Lys Arg Ile Thr Ala Thr Gln Ala Leu Glu 115 120 125 cat gag tat ttt cgc atg gaa cct ttg ccg gga cgc aac gct ctg gta 431 His Glu Tyr Phe Arg Met Glu Pro Leu Pro Gly Arg Asn Ala Leu Val 130 135 140 cca cca cag cct ggg gag aaa att gtg aac tac cca aca cga cca gtg 479 Pro Pro Gln Pro Gly Glu Lys Ile Val Asn Tyr Pro Thr Arg Pro Val 145 150 155 gac aca aat act gat att gaa gga aca atc agc ctc cag ccc tct caa 527 Asp Thr Asn Thr Asp Ile Glu Gly Thr Ile Ser Leu Gln Pro Ser Gln 160 165 170 175 ccg gta tca tct ggg aat tct gtg tct ggg gcc cta gcc ggt cct cat 575 Pro Val Ser Ser Gly Asn Ser Val Ser Gly Ala Leu Ala Gly Pro His 180 185 190 gta atg caa aat aga tcc atg cct cgg cca atg ccc atg gtt ggc gtg 623 Val Met Gln Asn Arg Ser Met Pro Arg Pro Met Pro Met Val Gly Val 195 200 205 caa cgc atg caa cct cca ggg atc cca cac tat ggt ctt gct tct cag 671 Gln Arg Met Gln Pro Pro Gly Ile Pro His Tyr Gly Leu Ala Ser Gln 210 215 220 gca gga atg ggt gga gta aat cct ggt ggc atc cca att cag cgg gga 719 Ala Gly Met Gly Gly Val Asn Pro Gly Gly Ile Pro Ile Gln Arg Gly 225 230 235 gtt cct gct cag gct cat caa cag cag cag atg aga agg aaa gac cct 767 Val Pro Ala Gln Ala His Gln Gln Gln Gln Met Arg Arg Lys Asp Pro 240 245 250 255 gga atg ggg atg act gga tat cct cca caa cag aaa tca agg cgc ttt 815 Gly Met Gly Met Thr Gly Tyr Pro Pro Gln Gln Lys Ser Arg Arg Phe 260 265 270 tgagagtccg ggtggatttg gagcctaagt gggaggacaa atacacattc caatcaaatt 875 agaggaaacc ttaaattaat cttccagtca gctgaaacga caccagtgga accaaatgat 935 ctgaccccat ttccaggatt gcatgtattt attaggagga atacacgaat gaagattcga 995 gtctagtgcc aaattattct aacatacctt catcatttgt tcctactaca ttccgacgtt 1055 atatgtttca actagtggaa gggtttctgc agtccaccca tgtggcacaa acatgattca 1115 tagcatgcca agcaacactt tactggtgtg taccaaggca atttctctat ttccaagcca 1175 aaaaaaaaaa aaaaaaaa 1193 <210> 56 <211> 271 <212> PRT <213> Avicennia marina <400> 56 Ala Pro Glu Leu Leu Leu Gly Ala Lys His Tyr Thr Ser Ala Val Asp 1 5 10 15 Met Trp Ala Val Gly Cys Ile Phe Ala Glu Leu Leu Thr Leu Lys Pro 20 25 30 Leu Phe Gln Gly Gln Glu Val Lys Gly Thr Ser Asn Pro Phe Gln Leu 35 40 45 Asp Gln Leu Asp Lys Ile Phe Lys Val Leu Gly His Pro Thr Gln Glu 50 55 60 Lys Trp Pro Thr Leu Ala Asn Leu Pro His Trp Gln Ser Asp Val Gln 65 70 75 80 Arg Ile Gln Gly Leu Lys Tyr Asp Asn Thr Gly Leu Tyr Asn Val Val 85 90 95 His Leu Ser Pro Lys Asn Pro Ala Tyr Asp Leu Leu Ser Lys Met Leu 100 105 110 Glu Tyr Asp Pro Arg Lys Arg Ile Thr Ala Thr Gln Ala Leu Glu His 115 120 125 Glu Tyr Phe Arg Met Glu Pro Leu Pro Gly Arg Asn Ala Leu Val Pro 130 135 140 Pro Gln Pro Gly Glu Lys Ile Val Asn Tyr Pro Thr Arg Pro Val Asp 145 150 155 160 Thr Asn Thr Asp Ile Glu Gly Thr Ile Ser Leu Gln Pro Ser Gln Pro 165 170 175 Val Ser Ser Gly Asn Ser Val Ser Gly Ala Leu Ala Gly Pro His Val 180 185 190 Met Gln Asn Arg Ser Met Pro Arg Pro Met Pro Met Val Gly Val Gln 195 200 205 Arg Met Gln Pro Pro Gly Ile Pro His Tyr Gly Leu Ala Ser Gln Ala 210 215 220 Gly Met Gly Gly Val Asn Pro Gly Gly Ile Pro Ile Gln Arg Gly Val 225 230 235 240 Pro Ala Gln Ala His Gln Gln Gln Gln Met Arg Arg Lys Asp Pro Gly 245 250 255 Met Gly Met Thr Gly Tyr Pro Pro Gln Gln Lys Ser Arg Arg Phe 260 265 270 <210> 57 <211> 1195 <212> DNA <213> Sueada japonica <220> <221> CDS <222> (116) .. (1195) <400> 57 gcaaaagtaa gagtgaaaga acacaaacca actttctatt ttcagctcaa atcaaattca 60 atagtggcaa aacaatagag ggcaaattct cattgcccaa ttcaaatttg gtaaa atg 118 Met 1 gct caa aag cat ttg aaa gaa ctt ctc aaa gaa gat caa gaa ccc ttt 166 Ala Gln Lys His Leu Lys Glu Leu Leu Lys Glu Asp Gln Glu Pro Phe 5 10 15 cat tta aag gat tac att gca act aaa aaa tgt caa ctt ttg aag aag 214 His Leu Lys Asp Tyr Ile Ala Thr Lys Lys Cys Gln Leu Leu Lys Lys 20 25 30 caa gaa tta gta gta ccc aaa tca aaa ctt caa ctc aaa aag cca aag 262 Gln Glu Leu Val Val Pro Lys Ser Lys Leu Gln Leu Lys Lys Pro Lys 35 40 45 cca aaa cca att tca aaa agc act tca gtt ttg tgc aaa aat gct tgc 310 Pro Lys Pro Ile Ser Lys Ser Thr Ser Val Leu Cys Lys Asn Ala Cys 50 55 60 65 ttt tta tct tta caa gaa tcc cct gac ctc aga aaa tcc ccc aaa cta 358 Phe Leu Ser Leu Gln Glu Ser Pro Asp Leu Arg Lys Ser Pro Lys Leu 70 75 80 ttt gat ttt cca cct tcc cct gtt tct aac aaa agc cca aac aga gta 406 Phe Asp Phe Pro Pro Ser Pro Val Ser Asn Lys Ser Pro Asn Arg Val 85 90 95 ttc ctc aat gtt cct gct aaa act gct gct ctt ctt ctt gaa gct gct 454 Phe Leu Asn Val Pro Ala Lys Thr Ala Ala Leu Leu Leu Glu Ala Ala 100 105 110 att cga att caa acc cac aaa tct aaa ccc aaa acc cag att aaa aat 502 Ile Arg Ile Gln Thr His Lys Ser Lys Pro Lys Thr Gln Ile Lys Asn 115 120 125 tcg ggt ttt ggg cta ttc ggg tca atg tta aag cga tta aat ctt cga 550 Ser Gly Phe Gly Leu Phe Gly Ser Met Leu Lys Arg Leu Asn Leu Arg 130 135 140 145 aat cgt acc caa aaa atc aag tca aaa aca gag gaa caa aac aga gga 598 Asn Arg Thr Gln Lys Ile Lys Ser Lys Thr Glu Glu Gln Asn Arg Gly 150 155 160 tgc tct gtt ttg agg agt gtt gaa gaa gaa aaa act acc acc att tct 646 Cys Ser Val Leu Arg Ser Val Glu Glu Glu Lys Thr Thr Thr Ile Ser 165 170 175 tct tct tca tct tca tct tct tca aca tca tcg tat tct tcg tgt tct 694 Ser Ser Ser Ser Ser Ser Ser Ser Ser Thr Ser Ser Tyr Ser Ser Cys Ser 180 185 190 tgc aat gag agg tta agt agt ttg gat ttg gag agt tct agc agt gga 742 Cys Asn Glu Arg Leu Ser Ser Leu Asp Leu Glu Ser Ser Ser Ser Gly 195 200 205 aga tca tta cat gat gaa gat gaa gat gaa gat gaa gat gat gaa ttt 790 Arg Ser Leu His Asp Glu Asp Glu Asp Glu Asp Glu Asp Asp Glu Phe 210 215 220 225 gag ttt aca aat gtt tta aga gaa aat aat aat gat gat aaa aat gga 838 Glu Phe Thr Asn Val Leu Arg Glu Asn Asn Asn Asp Asp Lys Asn Gly 230 235 240 ggt tat tat tca gga att tgc tta agt cct ttg agt cca ttt cgt ttt 886 Gly Tyr Tyr Ser Gly Ile Cys Leu Ser Pro Leu Ser Pro Phe Arg Phe 245 250 255 gct ctt cat aaa aac tct tct cct gaa cgt tgc tct cct gct aaa tcc 934 Ala Leu His Lys Asn Ser Ser Pro Glu Arg Cys Ser Pro Ala Lys Ser 260 265 270 cct gtt cgt tgc aaa ttt gag ggt aat gct aaa tat gaa caa gaa agc 982 Pro Val Arg Cys Lys Phe Glu Gly Asn Ala Lys Tyr Glu Gln Glu Ser 275 280 285 tta ata aag ttt gaa gac gaa gat gaa gaa gac aaa gag caa aat agc 1030 Leu Ile Lys Phe Glu Asp Glu Asp Glu Glu Asp Lys Glu Gln Asn Ser 290 295 300 305 cct gtt tcc gtg ctc gat cct cca ttc gag gat gat tac gat ggg cat 1078 Pro Val Ser Val Leu Asp Pro Pro Phe Glu Asp Asp Tyr Asp Gly His 310 315 320 gag gag gat agc tac gag gac atc gaa tgc agc tat gct ttt gta caa 1126 Glu Glu Asp Ser Tyr Glu Asp Ile Glu Cys Ser Tyr Ala Phe Val Gln 325 330 335 aga gca caa caa gag tta ttg cac aga ctt cac cgg ttc cag aag cta 1174 Arg Ala Gln Gln Glu Leu Leu His Arg Leu His Arg Phe Gln Lys Leu 340 345 350 gcg gag ttg gac cca att gaa 1195 Ala Glu Leu Asp Pro Ile Glu 355 360 <210> 58 <211> 360 <212> PRT <213> Sueada japonica <400> 58 Met Ala Gln Lys His Leu Lys Glu Leu Leu Lys Glu Asp Gln Glu Pro 1 5 10 15 Phe His Leu Lys Asp Tyr Ile Ala Thr Lys Lys Cys Gln Leu Leu Lys 20 25 30 Lys Gln Glu Leu Val Val Pro Lys Ser Lys Leu Gln Leu Lys Lys Pro 35 40 45 Lys Pro Lys Pro Ile Ser Lys Ser Thr Ser Val Leu Cys Lys Asn Ala 50 55 60 Cys Phe Leu Ser Leu Gln Glu Ser Pro Asp Leu Arg Lys Ser Pro Lys 65 70 75 80 Leu Phe Asp Phe Pro Pro Ser Pro Val Ser Asn Lys Ser Pro Asn Arg 85 90 95 Val Phe Leu Asn Val Pro Ala Lys Thr Ala Ala Leu Leu Leu Glu Ala 100 105 110 Ala Ile Arg Ile Gln Thr His Lys Ser Lys Pro Lys Thr Gln Ile Lys 115 120 125 Asn Ser Gly Phe Gly Leu Phe Gly Ser Met Leu Lys Arg Leu Asn Leu 130 135 140 Arg Asn Arg Thr Gln Lys Ile Lys Ser Lys Thr Glu Glu Gln Asn Arg 145 150 155 160 Gly Cys Ser Val Leu Arg Ser Val Glu Glu Glu Lys Thr Thr Thr Ile 165 170 175 Ser Ser Ser Ser Ser Ser Ser Ser Ser Thr Ser Ser Tyr Ser Ser Cys 180 185 190 Ser Cys Asn Glu Arg Leu Ser Ser Leu Asp Leu Glu Ser Ser Ser Ser 195 200 205 Gly Arg Ser Leu His Asp Glu Asp Glu Asp Glu Asp Glu Asp Asp Glu 210 215 220 Phe Glu Phe Thr Asn Val Leu Arg Glu Asn Asn Asn Asp Asp Lys Asn 225 230 235 240 Gly Gly Tyr Tyr Ser Gly Ile Cys Leu Ser Pro Leu Ser Pro Phe Arg 245 250 255 Phe Ala Leu His Lys Asn Ser Ser Pro Glu Arg Cys Ser Pro Ala Lys 260 265 270 Ser Pro Val Arg Cys Lys Phe Glu Gly Asn Ala Lys Tyr Glu Gln Glu 275 280 285 Ser Leu Ile Lys Phe Glu Asp Glu Asp Glu Glu Asp Lys Glu Gln Asn 290 295 300 Ser Pro Val Ser Val Leu Asp Pro Pro Phe Glu Asp Asp Tyr Asp Gly 305 310 315 320 His Glu Glu Asp Ser Tyr Glu Asp Ile Glu Cys Ser Tyr Ala Phe Val 325 330 335 Gln Arg Ala Gln Gln Glu Leu Leu His Arg Leu His Arg Phe Gln Lys 340 345 350 Leu Ala Glu Leu Asp Pro Ile Glu 355 360 <210> 59 <211> 1301 <212> DNA <213> Salsola komarovii <220> <221> CDS <222> (3) .. (815) <400> 59 gt gag gtt gac gat agc gtt aat agt cta cag gca gat gtt gac aac 47 Glu Val Asp Asp Ser Val Asn Ser Leu Gln Ala Asp Val Asp Asn 1 5 10 15 ctt tca att gag gaa cgc aga ttg gat gaa cag ata agg gaa atg caa 95 Leu Ser Ile Glu Glu Arg Arg Leu Asp Glu Gln Ile Arg Glu Met Gln 20 25 30 gaa aga ttg agg gaa atg agt gaa gat gat atc aat cag aag tgg ctt 143 Glu Arg Leu Arg Glu Met Ser Glu Asp Asp Ile Asn Gln Lys Trp Leu 35 40 45 ttt gta act gaa gaa gac ata aag ggt tta cct tgt ttt cag aat gaa 191 Phe Val Thr Glu Glu Asp Ile Lys Gly Leu Pro Cys Phe Gln Asn Glu 50 55 60 acc tta att gca att aaa gct cca cat gga aca act ttg gag gtt cca 239 Thr Leu Ile Ala Ile Lys Ala Pro His Gly Thr Thr Leu Glu Val Pro 65 70 75 gat cca gat gag gct gtc gat tat cct caa aga aga tac aag ata gtt 287 Asp Pro Asp Glu Ala Val Asp Tyr Pro Gln Arg Arg Tyr Lys Ile Val 80 85 90 95 ctt agg agc aca atg ggt cct att gat gta tat tta gtc agt caa ttt 335 Leu Arg Ser Thr Met Gly Pro Ile Asp Val Tyr Leu Val Ser Gln Phe 100 105 110 gaa gag aag ttt gag gag atc agt ggt gct gac ggt cca cta agt ata 383 Glu Glu Lys Phe Glu Glu Ile Ser Gly Ala Asp Gly Pro Leu Ser Ile 115 120 125 cca agt acc tca ggt gat gac aaa cac aca act gtt gca gct aag gaa 431 Pro Ser Thr Ser Gly Asp Asp Lys His Thr Thr Val Ala Ala Lys Glu 130 135 140 gaa agc aat ggc aat gag att gaa ata gaa gga caa ggg acc cat aga 479 Glu Ser Asn Gly Asn Glu Ile Glu Ile Glu Gly Gln Gly Thr His Arg 145 150 155 atc tgc tca gat tcc aac gct cag caa gac ttt gtg agt gga att atg 527 Ile Cys Ser Asp Ser Asn Ala Gln Gln Asp Phe Val Ser Gly Ile Met 160 165 170 175 aag ata gtg cct gaa gtt gat agt gat gca gat tac tgg ttg cta tcg 575 Lys Ile Val Pro Glu Val Asp Ser Asp Ala Asp Tyr Trp Leu Leu Ser 180 185 190 gat gct gat gtt agc att act gac atg tgg gga act gat tct gga gtt 623 Asp Ala Asp Val Ser Ile Thr Asp Met Trp Gly Thr Asp Ser Gly Val 195 200 205 gaa tgg aat gaa tta ggg act ata cat gaa gac tat gcc gtg gct aat 671 Glu Trp Asn Glu Leu Gly Thr Ile His Glu Asp Tyr Ala Val Ala Asn 210 215 220 gtt ggc act tca cag cca caa agt cca cca aca agt gca aca gaa gtg 719 Val Gly Thr Ser Gln Pro Gln Ser Pro Pro Thr Ser Ala Thr Glu Val 225 230 235 ctt cca gct aac atg aca agc agg aga ttg aca tgg agt ttt gag aga 767 Leu Pro Ala Asn Met Thr Ser Arg Arg Leu Thr Trp Ser Phe Glu Arg 240 245 250 255 att gcc aar att cat tca aat ggt cac tat tgc ttg gaa gtg agg ctc 815 Ile Ala Lys Ile His Ser Asn Gly His Tyr Cys Leu Glu Val Arg Leu 260 265 270 taactttcta ttattcatcc tgggatttgg gtacgaaagt ctgccttgaa gatgctgtaa 875 catgttgtgt attacaactg tgtgaatcta gtaagttggt agggtgagat tgttcctgat 935 cttattgcac agccggttgg gagagattga tcgctcaaca actgacaaaa ttggggcatg 995 ttaacggata gtatgcagtt gtaattttgt acatcacatt tgttgatttt agtcagtaca 1055 tcataactag ctcttcctat acttcttcaa ttgtcaactg gaatagattt ttagattaat 1115 tagatctctc tttgtatgga aatgtttcag ggtaacaagc cagaaattaa aatggtttta 1175 tgtgtaaaaa tatatactta aattgtttgt aggaagtttc tgatgggttg ttggatggct 1235 tttaacaact acatcgtata aggaaattcg tatcacaaat tcacaatgaa aaaaaaaaaa 1295 aaaaaa 1301 <210> 60 <211> 271 <212> PRT <213> Salsola komarovii <400> 60 Glu Val Asp Asp Ser Val Asn Ser Leu Gln Ala Asp Val Asp Asn Leu 1 5 10 15 Ser Ile Glu Glu Arg Arg Leu Asp Glu Gln Ile Arg Glu Met Gln Glu 20 25 30 Arg Leu Arg Glu Met Ser Glu Asp Asp Ile Asn Gln Lys Trp Leu Phe 35 40 45 Val Thr Glu Glu Asp Ile Lys Gly Leu Pro Cys Phe Gln Asn Glu Thr 50 55 60 Leu Ile Ala Ile Lys Ala Pro His Gly Thr Thr Leu Glu Val Pro Asp 65 70 75 80 Pro Asp Glu Ala Val Asp Tyr Pro Gln Arg Arg Tyr Lys Ile Val Leu 85 90 95 Arg Ser Thr Met Gly Pro Ile Asp Val Tyr Leu Val Ser Gln Phe Glu 100 105 110 Glu Lys Phe Glu Glu Ile Ser Gly Ala Asp Gly Pro Leu Ser Ile Pro 115 120 125 Ser Thr Ser Gly Asp Asp Lys His Thr Thr Val Ala Ala Lys Glu Glu 130 135 140 Ser Asn Gly Asn Glu Ile Glu Ile Glu Gly Gln Gly Thr His Arg Ile 145 150 155 160 Cys Ser Asp Ser Asn Ala Gln Gln Asp Phe Val Ser Gly Ile Met Lys 165 170 175 Ile Val Pro Glu Val Asp Ser Asp Ala Asp Tyr Trp Leu Leu Ser Asp 180 185 190 Ala Asp Val Ser Ile Thr Asp Met Trp Gly Thr Asp Ser Gly Val Glu 195 200 205 Trp Asn Glu Leu Gly Thr Ile His Glu Asp Tyr Ala Val Ala Asn Val 210 215 220 Gly Thr Ser Gln Pro Gln Ser Pro Pro
Thr Ser Ala Thr Glu Val Leu 225 230
235 240 Pro Ala Asn Met Thr Ser Arg Arg Leu
Thr Trp Ser Phe Glu Arg Ile 245
250 255 Ala Lys Ile His Ser Asn Gly His Tyr
Cys Leu Glu Val Arg Leu 260 265
270 <210> 61 <211> 1032 <212> DNA <213> Salsola komarovii <220> <221> CDS <222> (1). . (732) <400> 61 cca caa cga aga ccc gac ccg gtc ccg
aac ctt cac ggt cag ctt ttt 48 Pro Gln Arg Arg Pro Asp Pro Val Pro
Asn Leu His Gly Gln Leu Phe 15
10 15 caa cac cga aat cca cac cac cgt gac
ctc cac ccc tgc cgt agc ccg 96 Gln His Arg Asn Pro His His Arg Asp
Leu His Pro Cys Arg Ser Pro gca atg ggt cca ctc cct ccg cag act cat ctg cgc tgg tat tcc ctc 144 Ala Met Gly Pro Leu Pro Pro Gln Thr His Leu Arg Trp Tyr Ser Leu 35 40 45 tcg cgc tac tcc ccc gtg atc ggc ctc ggc gtc caa tgg aag ccc tcc 192 Ser Arg Tyr Ser Pro Val Ile Gly Leu Gly Val Gln Trp Lys Pro Ser 50 55 60 tcc acc tca gct gcc act ctt caa ctc agc atc gac aaa aag tgc ctc 240 Ser Thr Ser Ala Ala Thr Leu Gln Leu Ser Ile Asp Lys Lys Cys Leu 65 70 75 80 atc ttc caa ctc tcc cac tcc ccc gcc atc ccc gcc acc ctc cgc gac 288 Ile Phe Gln Leu Ser His Ser Pro Ala Ile Pro Ala Thr Leu Arg Asp 85 90 95 ctc ctc ctc gac gat cgc gtc acc ttc ttt ggt gtc cac aac ggc cgt 336 Leu Leu Leu Asp Asp Arg Val Thr Phe Phe Gly Val His Asn Gly Arg 100 105 110 gcc cgc gac ctc ctc caa ggg tcc cac cat gag ctc gac gtc aac aat 384 Ala Arg Asp Leu Leu Gln Gly Ser His His Glu Leu Asp Val Asn Asn 115 120 125 ctg gtt gat ctt gcc gag gag gaa aat ggt cat tac ttg aag tgg tcc 432 Leu Val Asp Leu Ala Glu Glu Glu Asn Gly His Tyr Leu Lys Trp Ser 130 135 140 atg gaa gac atg gct gaa gat gtg ttg ggc ttt tgt ggg gta cac aaa 480 Met Glu Asp Met Ala Glu Asp Val Leu Gly Phe Cys Gly Val His Lys 145 150 155 160 ccc agg aag gtt atg tta agt ggt tgg gat cag tat tgc ttg tct aat 528 Pro Arg Lys Val Met Leu Ser Gly Trp Asp Gln Tyr Cys Leu Ser Asn 165 170 175 gac cag gtt cag tat gct tgt gtt gat gct tac gtt tct ctt cgt ctt 576 Asp Gln Val Gln Tyr Ala Cys Val Asp Ala Tyr Val Ser Leu Arg Leu 180 185 190 gct cga gct tat ggg tac cac cgt ctc gat cac gat gat gat tat gat 624 Ala Arg Ala Tyr Gly Tyr His Arg Leu Asp His Asp Asp Asp Tyr Asp 195 200 205 gac cat gac gac gat gat aac gac cac acc gat gat gat tac gat gac 672 Asp His Asp Asp Asp Asp Asn Asp His Thr Asp Asp Asp Tyr Asp Asp 210 215 220 gtt tac gac cgc aat ata ggc tct gat gat gat ggt tat gat gcc gat 720 Val Tyr Asp Arg Asn Ile Gly Ser Asp Asp Asp Gly Tyr Asp Ala Asp 225 230 235 240 gat gat cga cga tgatcaattt ggactagact tcgttattgg aagggtccga 772 Asp Asp Arg Arg tcatcatgcc agtctaatta caaagagaca agaaataaaa atgatgatca aaaaaagaag 832 tcaatccata tacgtaattt tcattgcaat atcaattttg aggtgtttta ttattggcct 892 gtaataatag ttttatttaa taatagcact atagatctca tcctaacctt tacttattgg 952 gcttatgcgc tgtatgtcca ataaccaagt ttaatttatt tcatgatctg atgattactg 1012 caaaaaaaaa aaaaaaaaaa 1032 <210> 62 <211> 244 <212> PRT <213> Salsola komarovii <400> 62 Pro Gln Arg Arg Pro Asp Pro Val Pro Asn Leu His Gly Gln Leu Phe 1 5 10 15 Gln His Arg Asn Pro His His Arg Asp Leu His Pro Cys Arg Ser Pro 20 25 30 Ala Met Gly Pro Leu Pro Pro Gln Thr His Leu Arg Trp Tyr Ser Leu 35 40 45 Ser Arg Tyr Ser Pro Val Ile Gly Leu Gly Val Gln Trp Lys Pro Ser 50 55 60 Ser Thr Ser Ala Ala Thr Leu Gln Leu Ser Ile Asp Lys Lys Cys Leu 65 70 75 80 Ile Phe Gln Leu Ser His Ser Pro Ala Ile Pro Ala Thr Leu Arg Asp 85 90 95 Leu Leu Leu Asp Asp Arg Val Thr Phe Phe Gly Val His Asn Gly Arg 100 105 110 Ala Arg Asp Leu Leu Gln Gly Ser His His Glu Leu Asp Val Asn Asn 115 120 125 Leu Val Asp Leu Ala Glu Glu Glu Asn Gly His Tyr Leu Lys Trp Ser 130 135 140 Met Glu Asp Met Ala Glu Asp Val Leu Gly Phe Cys Gly Val His Lys 145 150 155 160 Pro Arg Lys Val Met Leu Ser Gly Trp Asp Gln Tyr Cys Leu Ser Asn 165 170 175 Asp Gln Val Gln Tyr Ala Cys Val Asp Ala Tyr Val Ser Leu Arg Leu 180 185 190 Ala Arg Ala Tyr Gly Tyr His Arg Leu Asp His Asp Asp Asp Tyr Asp 195 200 205 Asp His Asp Asp Asp Asp Asn Asp His Thr Asp Asp Asp Tyr Asp Asp 210 215 220 Val Tyr Asp Arg Asn Ile Gly Ser Asp Asp Asp Gly Tyr Asp Ala Asp 225 230 235 240 Asp Asp Arg Arg  <210> 63 <211> 1029 <212> DNA <213> Mesembryanthemum crystallinum <220> <221> CDS <222> (3) .. (824) <400> 63 ca cat atc agc cac atc cac tta att ccc cac agt ctt agt ctc tta 47 His Ile Ser His Ile His Leu Ile Pro His Ser Leu Ser Leu Leu 1 5 10 15 gac acc cat ctt agt ctt aag cct ctc atg gcc acc gcg gta ttc tca 95 Asp Thr His Leu Ser Leu Lys Pro Leu Met Ala Thr Ala Val Phe Ser 20 25 30 cct tct gcc ctt cta tcc acc tcc aca tcc acc tca aca acc cct ctt 143 Pro Ser Ala Leu Leu Ser Thr Ser Thr Ser Thr Ser Thr Thr Pro Leu 35 40 45 aaa gct ccc ccc ttg gcc tta acc aag acc cac gta acg atc cca tca 191 Lys Ala Pro Pro Leu Ala Leu Thr Lys Thr His Val Thr Ile Pro Ser 50 55 60 tca tca aag cca ccc cta acc aat tta act acc agt tta act gct gtc 239 Ser Ser Lys Pro Pro Leu Thr Asn Leu Thr Thr Ser Leu Thr Ala Val 65 70 75 gcc aca gct gct gcc ata atc ctg tcc aca acc cct cca tcg ttt gct 287 Ala Thr Ala Ala Ala Ile Ile Leu Ser Thr Thr Pro Pro Ser Phe Ala 80 85 90 95 gat gat ttg cag aca aat gca tac aac att tac tac ggc act gct gca 335 Asp Asp Leu Gln Thr Asn Ala Tyr Asn Ile Tyr Tyr Gly Thr Ala Ala 100 105 110 agt gca gcc aat tat gga ggc tac ggt ggc aat tcg aac aag aaa gat 383 Ser Ala Ala Asn Tyr Gly Gly Tyr Gly Gly Asn Ser Asn Lys Lys Asp 115 120 125 tca gct gag tac ata tat gac gtc cct gca ggt tgg aaa gag aga cta 431 Ser Ala Glu Tyr Ile Tyr Asp Val Pro Ala Gly Trp Lys Glu Arg Leu 130 135 140 gta tca aaa gtt gag aag ggt acc aat gga aca gat agt gag ttc ttc 479 Val Ser Lys Val Glu Lys Gly Thr Asn Gly Thr Asp Ser Glu Phe Phe 145 150 155 aac ccc aag aag aag aca gag cga gag tac ctt acc tac ctt gct ggt 527 Asn Pro Lys Lys Lys Thr Glu Arg Glu Tyr Leu Thr Tyr Leu Ala Gly 160 165 170 175 att agg caa cta ggt ccc aaa gaa gtg atc ctc aac aac tta gca ctc 575 Ile Arg Gln Leu Gly Pro Lys Glu Val Ile Leu Asn Asn Leu Ala Leu 180 185 190 tca gat gtg aac ctg caa gat caa att tcc agt gca gac tct gtg aca 623 Ser Asp Val Asn Leu Gln Asp Gln Ile Ser Ser Ala Asp Ser Val Thr 195 200 205 tca gaa gag agg aaa gat gac aag gga cag gtt tac tat gat tat gag 671 Ser Glu Glu Arg Lys Asp Asp Lys Gly Gln Val Tyr Tyr Asp Tyr Glu 210 215 220 att gct gga gct ggt tca cac agt ttg ata tcg gta aca tgt gcc agg 719 Ile Ala Gly Ala Gly Ser His Ser Leu Ile Ser Val Thr Cys Ala Arg 225 230 235 aac aag cta tat gcg cat ttt gtt agc gca cca aca ccc gaa tgg aat 767 Asn Lys Leu Tyr Ala His Phe Val Ser Ala Pro Thr Pro Glu Trp Asn 240 245 250 255 cgg gat caa gat atg ctg agg cac atc cac aac tca ttt aca aca gtc 815 Arg Asp Gln Asp Met Leu Arg His Ile His Asn Ser Phe Thr Thr Val 260 265 270 ggg tca ttc tagaaagtgt atatgataat catttataga gatgtcagag 864 Gly Ser Phe aggcatacat ttgaatgtac ttctgatgag ctggacttct tgatctatgt aacattgtaa 924 cgaaaattct ttctgggtta tcagaaacct agtgagtgct tgaaacttgc aatgagaaac 984 tcttcaataa acaatgactt gtatcaaaaa aaaaaaaaaa aaaaa 1029 <210> 64 <211> 274 <212> PRT <213> Mesembryanthemum crystallinum <400> 64 His Ile Ser His Ile His Leu Ile Pro His Ser Leu Ser Leu Leu Asp 1 5 10 15 Thr His Leu Ser Leu Lys Pro Leu Met Ala Thr Ala Val Phe Ser Pro 20 25 30 Ser Ala Leu Leu Ser Thr Ser Thr Ser Thr Ser Thr Thr Pro Leu Lys 35 40 45 Ala Pro Pro Leu Ala Leu Thr Lys Thr His Val Thr Ile Pro Ser Ser 50 55 60 Ser Lys Pro Pro Leu Thr Asn Leu Thr Thr Ser Leu Thr Ala Val Ala 65 70 75 80 Thr Ala Ala Ala Ile Ile Leu Ser Thr Thr Pro Pro Ser Phe Ala Asp 85 90 95 Asp Leu Gln Thr Asn Ala Tyr Asn Ile Tyr Tyr Gly Thr Ala Ala Ser 100 105 110 Ala Ala Asn Tyr Gly Gly Tyr Gly Gly Asn Ser Asn Lys Lys Asp Ser 115 120 125 Ala Glu Tyr Ile Tyr Asp Val Pro Ala Gly Trp Lys Glu Arg Leu Val 130 135 140 Ser Lys Val Glu Lys Gly Thr Asn Gly Thr Asp Ser Glu Phe Phe Asn 145 150 155 160 Pro Lys Lys Lys Thr Glu Arg Glu Tyr Leu Thr Tyr Leu Ala Gly Ile 165 170 175 Arg Gln Leu Gly Pro Lys Glu Val Ile Leu Asn Asn Leu Ala Leu Ser 180 185 190 Asp Val Asn Leu Gln Asp Gln Ile Ser Ser Ala Asp Ser Val Thr Ser 195 200 205 Glu Glu Arg Lys Asp Asp Lys Gly Gln Val Tyr Tyr Asp Tyr Glu Ile 210 215 220 Ala Gly Ala Gly Ser His Ser Leu Ile Ser Val Thr Cys Ala Arg Asn 225 230 235 240 Lys Leu Tyr Ala His Phe Val Ser Ala Pro Thr Pro Glu Trp Asn Arg 245 250 255 Asp Gln Asp Met Leu Arg His Ile His Asn Ser Phe Thr Thr Val Gly 260 265 270 Ser Phe  <210> 65 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Primer <400> 65 gctctgagaa ccgtctagac ttagatgaag gtg 33 <210> 66 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Primer <400> 66 tctctcgttc atctcgagct attacagctc 30} 

[Brief description of the drawings]

FIG. 1 shows the results of detecting the salt tolerance of Escherichia coli (SOLR) into which manga1 has been introduced. Detection was performed using colony formation as an index. Vector only (pBluescrip
t SK) was used. The assay was performed at two salt (NaCl) concentrations.

FIG. 2 shows the results of detecting the salt tolerance of Escherichia coli (SOLR) into which various partial sequences of mang1 have been introduced. Detection is
Colony formation was performed as an index. As a control, only the vector (pBluescript SK) was used. Two salts (NaCl)
The assay was performed at the concentration. Numerical values shown in parentheses indicate amino acid numbers, and “*” indicates manga1 cDNA including a coding region and a non-coding region.

FIG. 3 is a view showing the results of time-dependent measurement of the growth of yeast into which manga1 has been introduced under high salt conditions. Detection was performed using the cell concentration as an index. Vector alone (pYES2) was used as a control. The assay was performed at two salt (NaCl) concentrations.

FIG. 4 is a view showing the results of detecting the growth of cultivated tobacco cells into which man1 has been introduced under high salt conditions. Detection was performed using wet weight as an index. Vector alone (GUS) was used as a control. The assay was performed at three salt (NaCl) concentrations.

FIG. 5 is a diagram showing the results of detecting the growth of tobacco plants into which manga1 has been introduced under high salt conditions (150 mM NaCl). A and C are tobacco transfected with the vector alone (GUS) as a control, and B and D are mang1
2 shows a cigarette into which is introduced.

FIG. 6 shows the results of examining the heat stress resistance of Escherichia coli (SOLR) into which manga1 has been introduced. The heat stress resistance was evaluated using a growth curve at 40 ° C. culture as an index. SO containing only vector (pBluescript SK) as control
LR was used.

FIG. 7 shows the results of examining the osmotic resistance of Escherichia coli (SOLR) into which man1 has been introduced. Osmotic pressure resistance is 800
The growth on 2YT agar medium containing mM sorbitol was evaluated as an index. Vector only (pBlues
SOL) was used.

FIG. 8 shows the results of examining the freeze stress resistance of Escherichia coli (SOLR) into which manga1 has been introduced. Freezing stress resistance was evaluated by the growth of the freeze-thawed bacterial cells on a 2YT agar medium. Vector only (pBlu
escript SK) was used.

FIG. 9 shows an area (ma
ng1 core) and the nucleotide sequence and amino acid sequence of the clone into which the mutation was introduced. Inversion characters in the base sequence and the amino acid sequence indicate the place where the mutation occurred.

FIG. 10 shows a region considered to be the minimum functional region of manga1 and a clone obtained by introducing a mutation into the region at 85 mM and 350 mM.
M shows the results of growth on an agar medium containing 500 mM NaCl. Vector only (pBluescript
SK) was used.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (reference) C12P 21/02 C12P 21/08 // C12N 1/21 C12N 15/00 ZNAA C12P 21/08 5/00 CF term ( Reference) 2B030 AA02 AA03 AB03 AD04 CA06 CA17 CA19 CB02 CD02 CD03 CD07 CD09 CD10 4B024 AA08 AA11 BA53 BA80 CA04 DA01 DA06 DA12 GA11 GA14 GA17 HA01 4B064 AG01 AG27 CA01 CA02 CA20 CC24 DA13 DA20 4B065 AA26A ABA ABA XA BA08 CA24 CA25 CA46 CA53 4H045 AA10 AA11 AA20 BA10 BA20 BA21 CA30 DA75 DA76 EA05 FA72 FA73 FA74 HA04 HA05

Claims (125)

[Claims] 1. A method for screening a DNA encoding a protein having an activity to improve environmental stress tolerance, comprising introducing a cDNA library-derived candidate cDNA into a host cell, and transforming the obtained transformed cell into a host cell. A screening method characterized by culturing under environmental conditions that cannot grow in a specific manner, selecting a clone growing after culturing, and isolating a candidate cDNA introduced from the selected clone. 2. A method for screening a DNA encoding a protein having an activity for improving environmental stress tolerance, comprising introducing a candidate cDNA derived from a cDNA library into a host cell, and transforming the obtained transformed cell into a host cell. Culturing under environmental conditions where growth is not possible, selecting clones growing after culturing, isolating candidate cDNAs introduced from the selected clones, isolated candidate cDNAs
A screening method comprising: repeating a step of introducing a random mutation into a host cell, introducing the mutant cDNA into a host cell, and selecting the cDNA under severer conditions than the selection conditions of the cDNA before the mutation one or more times. 3. The method according to claim 1, wherein the environmental stress is one or more stresses selected from chemical stress, high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress, radiation stress, and osmotic stress. The screening method according to claim 1 or 2, wherein 4. The screening method according to claim 3, wherein the chemical substance stress is a salt stress. 5. The screening method according to claim 1, wherein the host cell is Escherichia coli. 6. The screening method according to claim 5, wherein the Escherichia coli is a SOLR strain. 7. The screening method according to claim 1, wherein the environmental condition under which the host cell cannot substantially grow is a condition of a salt concentration of 350 mM or more. 8. A DN encoding a protein having an activity for improving environmental stress tolerance, which is obtained by the screening method according to any one of claims 1 to 7.
A. 9. The method according to claim 9, wherein the environmental stress is one or more stresses selected from chemical stress, high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress, radiation stress and osmotic stress. A DNA encoding a protein having an activity for improving environmental stress tolerance according to claim 8. 10. The DNA encoding a protein having an activity for improving environmental stress tolerance according to claim 9, wherein the chemical substance stress is salt stress. 11. The DNA encoding a protein having an activity of improving environmental stress tolerance according to claim 8, wherein the protein having an activity of improving environmental stress tolerance is a plant-derived protein. 12. The plant, wherein the plant is Bruguiera sexang
la), Hirugimasashi (Avicennia marina), turkey
The DNA encoding a protein having an activity to improve environmental stress tolerance according to claim 11, which is selected from the group consisting of (Sueada japonica), okajiki (Salsolakomarovii) and ice plant (Mesembryanthemum crystallinum). 13. A DNA encoding a protein according to any one of the following (a) to (c): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 2; (b) a protein having a homology of 70 to the amino acid sequence shown in SEQ ID NO: 2
% Or more and having at least an activity of improving salt stress tolerance. (C) An amino acid sequence represented by SEQ ID NO: 2 in which one or several amino acids are deleted, substituted or added. , And at least a protein having activity to improve salt stress tolerance 14. A DNA comprising the nucleotide sequence shown in SEQ ID NO: 1, its complementary sequence, or a part or all of these sequences. 15. A DNA that hybridizes with the DNA according to claim 14 under stringent conditions and encodes a protein having at least an activity of improving salt stress tolerance. 16. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 4; (b) an amino acid sequence shown in SEQ ID NO: 4 consisting of an amino acid sequence in which one or several amino acids are deleted, substituted or added, and at least a salt Protein having activity to improve stress tolerance 17. A DNA comprising the nucleotide sequence shown in SEQ ID NO: 3, its complementary sequence, or a part or all of these sequences. 18. A DNA that hybridizes with the DNA according to claim 17 under stringent conditions and encodes a protein having at least an activity of improving salt stress tolerance. 19. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 6; (b) an amino acid sequence shown in SEQ ID NO: 6 consisting of an amino acid sequence in which one or several amino acids are deleted, substituted or added, and at least a salt Protein having activity to improve stress tolerance 20. A DNA comprising the nucleotide sequence shown in SEQ ID NO: 5, its complementary sequence, or a part or all of these sequences. 21. D constituting the gene according to claim 20.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 22. A DNA encoding the protein of any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 8; (b) an amino acid sequence shown in SEQ ID NO: 8 consisting of an amino acid sequence in which one or several amino acids are deleted, substituted or added, and at least a salt Protein having activity to improve stress tolerance 23. A DNA comprising the base sequence shown in SEQ ID NO: 7, a complementary sequence thereof, or a part or all of these sequences. 24. D constituting the gene according to claim 23.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 25. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 10 (b) in the amino acid sequence shown in SEQ ID NO: 10
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 26. A DNA comprising the base sequence shown in SEQ ID NO: 9, its complementary sequence, or a part or all of these sequences. 27. D constituting the gene according to claim 26.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 28. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 12; (b) in the amino acid sequence shown in SEQ ID NO: 12, 1
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 29. A DNA comprising the nucleotide sequence shown in SEQ ID NO: 11, its complementary sequence, or a part or all of these sequences. 30. D constituting the gene according to claim 29.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 31. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 14;
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 32. A DNA comprising the base sequence shown in SEQ ID NO: 13, a complementary sequence thereof, or a part or all of these sequences. 33. D constituting the gene according to claim 32.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 34. A DNA encoding the protein of any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 16; (b) in the amino acid sequence shown in SEQ ID NO: 16,
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 35. A DNA comprising the base sequence shown in SEQ ID NO: 15, a complementary sequence thereof, or a part or all of these sequences. 36. D constituting the gene according to claim 35.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 37. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 18; (b) in the amino acid sequence shown in SEQ ID NO: 18, 1
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 38. A DNA comprising the base sequence shown in SEQ ID NO: 17, a complementary sequence thereof, or a part or all of these sequences. 39. D constituting the gene according to claim 38.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 40. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 20 (b) in the amino acid sequence shown in SEQ ID NO: 20
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 41. A DNA comprising the base sequence shown in SEQ ID NO: 19, its complementary sequence, or a part or all of these sequences. 42. D constituting the gene according to claim 41.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 43. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 22; (b) in the amino acid sequence shown in SEQ ID NO: 22
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 44. A DNA comprising the base sequence shown in SEQ ID NO: 21 or a sequence complementary thereto, or a part or all of these sequences. (45) D constituting the gene according to (44);
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 46. A DNA encoding the protein of any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 24; (b) in the amino acid sequence shown in SEQ ID NO: 24,
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 47. A DNA comprising the nucleotide sequence shown in SEQ ID NO: 23, its complementary sequence, or a part or all of these sequences. 48. D constituting the gene according to claim 47.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 49. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 26; (b) in the amino acid sequence shown in SEQ ID NO: 26,
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 50. A DNA comprising the nucleotide sequence shown in SEQ ID NO: 25, its complementary sequence, or a part or all of these sequences. 51. D constituting the gene according to claim 50.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 52. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 28; (b) in the amino acid sequence shown in SEQ ID NO: 28,
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 53. A DNA comprising the nucleotide sequence of SEQ ID NO: 27, its complementary sequence, or a part or all of these sequences. 54. D constituting the gene according to claim 53.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 55. A DNA encoding the protein of any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 30; (b) in the amino acid sequence shown in SEQ ID NO: 30
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 56. A DNA comprising the base sequence shown in SEQ ID NO: 29, its complementary sequence, or a part or all of these sequences. 57. D constituting the gene according to claim 56.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 58. A DNA encoding the protein of any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 32; (b) in the amino acid sequence shown in SEQ ID NO: 32,
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 59. A DNA comprising the base sequence shown in SEQ ID NO: 31, a complementary sequence thereof, or a part or all of these sequences. 60. D constituting the gene according to claim 59.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 61. A DNA encoding a protein according to any one of the following (a) and (b): (a) a protein consisting of the amino acid sequence shown in SEQ ID NO: 34; (b) in the amino acid sequence shown in SEQ ID NO: 34,
Or a protein comprising an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance 62. A DNA comprising the nucleotide sequence shown in SEQ ID NO: 33, its complementary sequence, or a part or all of these sequences. 63. D constituting the gene according to claim 62.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 64. A DNA encoding a protein according to any one of the following (a) and (b): (a) SEQ ID NOs: 36, 38, 40, 42, 44, 46, 4
8, 50, 52, 54, 56, 58, 60, 62 or 6
(B) SEQ ID NOs: 36, 38, 40, 42, 44, 46, 4
8, 50, 52, 54, 56, 58, 60, 62 or 6
A protein comprising an amino acid sequence in which one or several amino acids have been deleted, substituted or added in the amino acid sequence shown in 4, and having at least an activity of improving salt stress tolerance 65. SEQ ID NOS: 35, 37, 39, 41, 4
3,45,47,49,51,53,55,57,5
A DNA comprising the base sequence shown in SEQ ID NO: 9, 61 or 63 or a complementary sequence thereof or a part or all of these sequences
A. 66. D constituting the gene according to claim 65.
Hybridizes with NA under stringent conditions,
And a DNA encoding a protein having at least a salt stress tolerance improving activity. 67. The DN according to any one of claims 8 to 66.
A method for improving environmental stress resistance, wherein A is used. 68. The environmental stress is a chemical stress,
68. The improvement of environmental stress tolerance according to claim 67, wherein the stress is one or more stress selected from high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress, radiation stress, and osmotic stress. Method. 69. The method for improving environmental stress tolerance according to claim 68, wherein the chemical substance stress is salt stress. 70. A protein consisting of the amino acid sequence shown in SEQ ID NO: 2. 71. A protein consisting of an amino acid sequence having a homology of 70% or more with the amino acid sequence shown in SEQ ID NO: 2 and having at least a salt stress tolerance improving activity. 72. A protein comprising the amino acid sequence of SEQ ID NO: 2 with one or several amino acids deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 73. A protein consisting of the amino acid sequence shown in SEQ ID NO: 4. 74. A protein comprising the amino acid sequence shown in SEQ ID NO: 4 with one or several amino acids deleted, substituted or added, and having at least a salt stress tolerance improving activity. 75. A protein consisting of the amino acid sequence shown in SEQ ID NO: 6. 76. A protein comprising the amino acid sequence of SEQ ID NO: 6 with one or several amino acids deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 77. A protein consisting of the amino acid sequence shown in SEQ ID NO: 8. 78. A protein comprising the amino acid sequence of SEQ ID NO: 8 with one or several amino acids deleted, substituted or added, and having at least an activity of improving salt stress resistance. 79. A protein consisting of the amino acid sequence shown in SEQ ID NO: 10. 80. A protein comprising the amino acid sequence of SEQ ID NO: 10 with one or several amino acids deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 81. A protein consisting of the amino acid sequence shown in SEQ ID NO: 12. 82. A protein comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 83. A protein consisting of the amino acid sequence shown in SEQ ID NO: 14. 84. A protein comprising the amino acid sequence of SEQ ID NO: 14 with one or several amino acids deleted, substituted or added and having at least an activity of improving salt stress tolerance. 85. A protein consisting of the amino acid sequence shown in SEQ ID NO: 16. 86. A protein consisting of the amino acid sequence shown in SEQ ID NO: 16 in which one or several amino acids have been deleted, substituted or added, and having at least a salt stress tolerance improving activity. 87. A protein consisting of the amino acid sequence shown in SEQ ID NO: 18. 88. A protein comprising the amino acid sequence of SEQ ID NO: 18 in which one or several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 89. A protein comprising the amino acid sequence shown in SEQ ID NO: 20. 90. A protein comprising the amino acid sequence of SEQ ID NO: 20 with one or several amino acids deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 91. A protein consisting of the amino acid sequence shown in SEQ ID NO: 22. 92. A protein comprising the amino acid sequence of SEQ ID NO: 22 with one or several amino acids deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 93. A protein consisting of the amino acid sequence shown in SEQ ID NO: 24. 94. A protein consisting of the amino acid sequence of SEQ ID NO: 24 with one or several amino acids deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 95. A protein consisting of the amino acid sequence shown in SEQ ID NO: 26. 96. A protein consisting of the amino acid sequence of SEQ ID NO: 26 in which one or several amino acids have been deleted, substituted or added, and having at least a salt stress tolerance improving activity. 97. A protein consisting of the amino acid sequence shown in SEQ ID NO: 28. 98. A protein consisting of the amino acid sequence of SEQ ID NO: 28 in which one or several amino acids have been deleted, substituted or added, and having at least a salt stress tolerance improving activity. 99. A protein comprising the amino acid sequence shown in SEQ ID NO: 30. 100. A protein comprising the amino acid sequence of SEQ ID NO: 30 with one or several amino acids deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 101. A protein comprising the amino acid sequence represented by SEQ ID NO: 32. 102. A protein consisting of the amino acid sequence of SEQ ID NO: 32 in which one or several amino acids have been deleted, substituted or added, and having at least a salt stress tolerance improving activity. 103. A protein consisting of the amino acid sequence shown in SEQ ID NO: 34. 104. A protein consisting of the amino acid sequence of SEQ ID NO: 34 with one or several amino acids deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 105. SEQ ID NOs: 36, 38, 40, 42,
44, 46, 48, 50, 52, 54, 56, 58, 6
A protein consisting of the amino acid sequence represented by 0, 62 or 64. 106. SEQ ID NOs: 36, 38, 40, 42,
44, 46, 48, 50, 52, 54, 56, 58, 6
In the amino acid sequence represented by 0, 62 or 64, 1
Alternatively, a protein consisting of an amino acid sequence in which several amino acids have been deleted, substituted or added, and having at least an activity of improving salt stress tolerance. 107. An antibody that specifically binds to the protein according to any one of claims 70 to 72. 108. An antibody that specifically binds to the protein of any one of claims 73 to 104. An antibody which specifically binds to the protein according to claim 105 or 106. 110. The antibody according to any one of claims 107 to 109, wherein the antibody is a monoclonal antibody. 111. A vector comprising a DNA encoding the protein having the activity of improving environmental stress tolerance according to any one of claims 8 to 12. A vector comprising the DNA according to any one of claims 13 to 15. A vector comprising the DNA according to any one of claims 16 to 63. A vector comprising the DNA according to any one of claims 64 to 66. 115. A transformed cell obtained by introducing the vector according to any one of claims 111 to 114 into a host cell. 116. The transformed cell according to claim 115, wherein the host cell is a plant cell. 117. A protein having an activity for improving environmental stress tolerance, which comprises culturing the transformed cell according to claim 115 or 116, and recovering the recombinant protein from the supernatant of the transformed cell or a culture solution thereof. Manufacturing method. 118. A DNA obtained by introducing a DNA encoding the protein having an activity of improving environmental stress tolerance according to any one of claims 8 to 12 into a plant cell, and allowing the plant cell to divide, proliferate, and redifferentiate. A transgenic plant characterized by being obtained. 119. A transgenic plant obtained by introducing the DNA according to any one of claims 13 to 15 into a plant cell and causing the plant cell to divide, proliferate, and redifferentiate. 120. A transgenic plant obtained by introducing the DNA according to any one of claims 16 to 63 into a plant cell and causing the plant cell to divide, proliferate, and redifferentiate. 121. A transgenic plant obtained by introducing the DNA according to any one of claims 64 to 66 into a plant cell, and causing the plant cell to divide, proliferate, and redifferentiate. 122. A vector according to any of claims 111 to 114, which is introduced into a plant cell to divide the plant cell.
A transgenic plant, which is obtained by performing growth and regeneration. 123. The environmental stress is one or more stresses selected from chemical stress, high temperature stress, low temperature stress, freezing stress, drying stress, ozone stress, ultraviolet stress, radiation stress, and osmotic stress. A transgenic plant according to any of claims 118 to 122. 124. The transgenic plant according to claim 123, wherein the chemical stress is salt stress. 125. A propagation material derived from the transgenic plant according to any one of claims 118 to 122.