CN110157746A - A kind of method of Microbe synthesis auximone - Google Patents

Abstract

The invention discloses a kind of methods of Microbe synthesis auximone, it is characterised in that following microorganism knocks out TrpR, dkgB, yeaE, yqhC, yqhD, dkgA, yahK and yjgB gene, height expression aroG in microorganism^fbr、pheA^fbr, KDC, ALDH and ARO8 gene, the purpose project microorganism of building can realize glucose fermentation synthesize phenylacetic acid.Further replace pheA^fbrGene is tyrA^fbrGene, so that the objective microbe modified through this can realize that glucose fermentation synthesizes p-hydroxyphenylaceticacid.Or replacement pheA^fbrGene is TrpE^fbr serA^fbrWith trpABCD gene so that through this modify objective microbe can realize glucose fermentation synthesis of indole acetic acid.The microorganism that the present invention is transformed can be realized by the microorganism conversion of glucose to phenylacetic acid, p-hydroxyphenylaceticacid and heteroauxin, provide a connecting way for the above-mentioned auximone of large-scale biosynthesis.

Description

A kind of method of Microbe synthesis auximone

Technical field

The invention belongs to microbial metabolism engineering fields, and in particular to transformation microbial metabolism approach is directly closed from monosaccharide At the method for auximone.

Background technique

Auximone is one of pheromones mostly important in plant, is endogenous growth generally existing in plant Element, it is most important for the growth and development of plant, have promote cell division, vascular bundle differentiation, stem elongation, apical dominance, Germination, lateral root and Adventitious root initiation, nodule formation, ovary wall growth, seed and fruit growth are beared fruit, raw in agricultural It has broad application prospects in production and economic status.These auximones include heteroauxin, phenylacetic acid, para hydroxybenzene second Acid etc..

The auximone being commercialized at present is to be synthesized by chemical method.Chemical method synthetic auxin need to be related to more A reaction step causes synthesis cost higher.By biotechnological method, realize that microbial fermentation monosaccharide synthesizes various auxin, Cost can be effectively reduced.

Summary of the invention

The purpose of the present invention is to provide a kind of methods of synthetic auxin in microbial body, construct in microorganism One is transformed into the metabolic pathway of auximone by monosaccharide.

To achieve the above object, the present invention provides Escherichia coli and Pichia pastoris synthetic auxin phenylacetic acid, The method of para hydroxybenzene acetaldehyde and heteroauxin, key step are as follows:

(1) gene constructed bacterial strain GDY is knocked out

It is set out with Escherichia coli MG1655 bacterial strain, successively knocks out the TrpR gene and seven aldehyde reductase genes in the bacterial strain (dkgB, yeaE, yqhC, yqhD, dkgA, yahK and yjgB), the Strain Designation for knocking out above-mentioned 8 genes is GDY.

(2) building produces phenylacetic acid engineering bacteria GDY1

Future home-brewed saccharomycete KDC and ARO8 gene and ALDH gene from Escherichia coli, recombination arrive carrier On pET28a (+), carrier pDY001 is obtained.By the aroG from Escherichia coli^fbrGene and pheA^fbrCarrier is arrived in gene, recombination On pBBR1MCS1, carrier pDY002 is obtained.Carrier pDY001 and pDY002 are transferred in Escherichia coli GDY, realize above-mentioned 5 A large amount expression of a gene in Escherichia coli, obtains colibacillus engineering strain GDY1.Expression releases feedback inhibition aroG^fbrGene and pheA^fbrGene is used to reinforce the synthesis of Escherichia coli phenylalanine intracellular.Expression ARO8 is for being catalyzed phenylpropyl alcohol Propylhomoserin converts phenylpyruvic acid, and expressing K DC is phenylacetaldehyde for being catalyzed phenylpyruvic acid decarboxylation, and expression ALDH is for being then catalyzed benzene second Oxidation of aldehydes formation phenylacetic acid.The GDY1 engineering bacteria of above-mentioned building effectively can convert phenylacetic acid for monosaccharide.

(3) building produces p-hydroxyphenylaceticacid engineering bacteria GDY2

By the aroG from Escherichia coli^fbrGene and tyrA^fbrGene recombinates onto carrier pBBR1MCS1, obtains carrier pDY003.It by carrier pDY001 that pDY003 and front are built while being transferred in Escherichia coli GDY, realizes aroG^fbr、 tyrA^fbr, KDC, ARO8 and ALDH5 genes in Escherichia coli a large amount expression, obtain colibacillus engineering strain GDY2. Expression releases the aroG of feedback inhibition^fbrGene and tyrA^fbrGene is used to reinforce the synthesis of Escherichia coli tyrosine intracellular.Expression ARO8 converts Single-chip microcomputer for catalytic tyrosine, and expressing K DC is to hydroxyl for being catalyzed Single-chip microcomputer decarboxylation Base phenylacetaldehyde, expression ALDH form p-hydroxyphenylaceticacid for being then catalyzed para hydroxybenzene oxidation of acetaldehyde.The GDY2 of above-mentioned building Engineering bacteria effectively can convert p-hydroxyphenylaceticacid for monosaccharide.

(4) building produces heteroauxin engineering bacteria GDY3

By the aroG from Escherichia coli^fbr、TrpE^fbrAnd serA^fbrGene is recombinated onto carrier pBBR1MCS1, is obtained Carrier pDY004.In trpABCD gene (including tetra- genes of trpA, trpB, trpC and trpD) recombination to carrier pDY001, obtain To carrier pDY005.Carrier pDY004 and pDY005 is transferred in Escherichia coli GDY simultaneously, realizes aroG^fbr、TrpE^fbr、 serA^fbr, trpABCD, KDC, ARO8 and ALDH7 genes in Escherichia coli a large amount expression, obtain colibacillus engineering Strain GDY3.Expression releases the aroG of feedback inhibition^fbr、TrpE^fbrAnd serA^fbrGene and trpABCD gene are for reinforcing large intestine bar The synthesis of tryptophan in mycetocyte.Expression ARO8 turns to indolepyruvic acid for being catalyzed tryptophan transfer, and expressing K DC is used for catalyzing indole Pyruvate decarboxylation is indoleacetaldehyde, and expression ALDH forms heteroauxin for subsequent catalyzing indole oxidation of acetaldehyde.Above-mentioned building GDY3 engineering bacteria effectively can convert heteroauxin for monosaccharide.

(5) building produces auximone engineering bacteria GDY4

By the ALDH gene from Escherichia coli, recombination arrives carrier pGAPZa, obtains carrier pDY006.Future home-brewed ferment KDC the and ARO8 gene of female bacterium, carrier pGAPZa is arrived in recombination respectively, obtains carrier pDY14 and pDY15.By pDY14 and pDY15 Respectively with BamHI and BglII double digestion, obtain KDC gene expression frame (comprising GAP promoter, KDC gene and AOX1TT) and ARO8 gene expression frame (including GAP promoter, ARO8 gene and AOX1TT).By KDC gene expression frame and ARO8 gene expression Frame is recombinated respectively to pAO815 carrier, obtains carrier pDY007 and pDY008.Carrier pDY006, pDY007 and pDY008 are distinguished It after linearisation, successively imports in Pichia pastoris GS115, realizes the constitutive expression of ARO8, ALDH and KDC gene, the engineering bacteria It is named as GDY4.Expression ARO8 turns to phenylpyruvic acid, ethyl-para-hydroxyphenyl ketone for being catalyzed phenylalanine, tyrosine and tryptophan transfer And indolepyruvic acid.Expressing K DC is separately converted to benzene second for being catalyzed phenylpyruvic acid, Single-chip microcomputer and indolepyruvic acid Aldehyde, para hydroxybenzene acetaldehyde and indoleacetaldehyde.Expression ALDH turns respectively for being catalyzed phenylacetaldehyde, para hydroxybenzene acetaldehyde and indoleacetaldehyde Turn to phenylacetic acid, p-hydroxyphenylaceticacid and heteroauxin.

It is an advantage of the invention that the metabolic pathway that foreign gene constructs a synthetic auxin is introduced in microorganism, It is final to realize using monosaccharide as the purpose of Material synthesis auximone, it does not need through excessive chemosynthesis reaction, reductionization The release for learning noxious material in synthesis process, decreases synthesis cost.Simultaneously as the microorganism speed of growth is fast, heredity behaviour Make technology maturation, and fermentation process not can cause environmental pollution destruction.It is micro- that Pichi strain of the invention can be used as agricultural Biology, directly using the nutriment synthetic auxin in ambient soil, to promote the growth of plant.

Detailed description of the invention

Fig. 1 colibacillus engineering GDY1 tunning GC-MS testing result.No. 1 peak represents internal standard benzoic acid, No. 2 peaks Represent product phenylacetic acid.

Fig. 2 colibacillus engineering GDY2 tunning GC-MS testing result.No. 1 peak represents internal standard benzoic acid, No. 2 peaks By-product phenylacetic acid is represented, No. 3 peaks represent product p-hydroxyphenylaceticacid.

Fig. 3 colibacillus engineering GDY3 tunning GC-MS testing result.No. 1 peak represents internal standard benzoic acid, No. 2 peaks By-product phenylacetic acid is represented, No. 3 peaks represent by-product p-hydroxyphenylaceticacid, and No. 4 peaks represent product heteroauxin.

Fig. 4 Pichia yeast engineering GDY4 tunning GC-MS testing result.No. 1 peak represents internal standard benzoic acid, No. 2 peaks Product heteroauxin is represented, No. 3 peaks represent p-hydroxyphenylaceticacid, and No. 4 peaks represent heteroauxin.

Specific embodiment

Following embodiment should not be construed as limiting the invention for further illustrating the present invention.

The building of embodiment 1 produces phenylacetic acid colibacillus engineering

1. the building of gene knock-out bacterial strain GDY

Using pKD4 plasmid as template, PCR amplification target practice segment, the segment includes (kan^rSequence and with purpose base to be knocked out Because of the homology arm of both ends complementation)；The target practice segment electricity of amplification is gone in the Escherichia coli with pKD46 plasmid.Utilize pKD46 The Red homologous recombination system having on plasmid, recombinates target gene to be knocked out with target practice segment, target gene is by resistance Gene replaces.Temperature-sensitive plasmid pCP20 is then imported into bacterial strain, is dyed using the FLP flippase removal purpose bacterial strain on the plasmid Resistant gene on body.Successively knocked out using the above method TrpR, dkgB in Escherichia coli MG1655 bacterial strain, yeaE, yqhC, YqhD, dkgA, yahK and yjgB gene obtain Escherichia coli DGY.The TrpR gene has core shown in SEQ ID NO:25 Nucleotide sequence coding；The TrpR gene has amino acid sequence shown in SEQ ID NO:26；The dkgB gene has SEQ It is nucleotide sequence coded shown in ID NO:27；The ALDH gene has amino acid sequence shown in SEQ ID NO:28；Institute YeaE gene is stated with nucleotide sequence coded shown in SEQ ID NO:29；The yeaE has shown in SEQ ID NO:30 Amino acid sequence；The yqhC gene has nucleotide sequence coded shown in SEQ ID NO:31；The yqhC gene has Amino acid sequence shown in SEQ ID NO:32；There is the yqhD gene nucleotide sequence shown in SEQ ID NO:33 to compile Code；The yqhD gene has amino acid sequence shown in SEQ ID NO:34；The dkgA gene has SEQ ID NO:35 Shown in it is nucleotide sequence coded；The dkgA gene has amino acid sequence shown in SEQ ID NO:36；The yahK base It is nucleotide sequence coded shown in SEQ ID NO:37 because having；The yahK gene has amino shown in SEQ ID NO:38 Acid sequence；The yjgB gene has nucleotide sequence coded shown in SEQ ID NO:39；The yjgB gene has SEQ Amino acid sequence shown in ID NO:40；Knock out the primer of gene are as follows: TrpR-F: atggcccaacaatcaccctattcagcagcgatggcagaacaggtgtaggctggagctgcttc

TrpR-R:tcaatcgcttttcagcaacacctcttccagccactggcgcacatatgaatatcctcctta

DkgB-F:atggctatccctgcatttggtttaggtactttccgtctgagtgtaggctggagctgcttc

DkgB-R:ttaatcccattcaggagccagaccttccgggctaaccaggcggcatatgaatatcctcctta

yeaE-F:atgcaacaaaaaatgattcaatttagtggcgatgtctcactggtgtaggctggagctgcttc

yeaE-R:tcacaccatatccagcgcagtttttccttttggtgccggatatcatatgaatatcctcctta

yqhC-F:atgctacaaaattgcgcacaatcaaattgccgcattattcctaagtgtaggctggagctgct tc

yqhC-R:ttaattcccctgcatcgcccgcattcttgccgcatcttcccccatatgaatatcctcctta

yqhD-F:atgaacaactttaatctgcacaccccaacccgcattctgttgtgtaggctggagctgcttc

yqhD-R:ttagcgggcggcttcgtatatacggcggctgacatccaacgcatatgaatatcctcctta

DkgA-F:atggctaatccaaccgttattaagctacaggatggcaatgtcgtgtagg ctggagctgcttc

DkgA-R:ttagccgccgaactggtcaggatcgggaccgagacgcttgccatatgaa tatcctcctta

yahK-F:atgaagatcaaagctgttggtgcatattccgctaaacaaccgtgtaggctggagctgcttc

yahK-R:tcagtctgttagtgtgcgattatcgataacaaaacgatattcatatgaatatcctcctta

yjgB-F:atgtcgatgataaaaagctatgccgcaaaagaagcgggcggcgtgtaggctggagctgcttc

yjgB-R:tcaaaaatcggctttcaacaccacgcggtaacgcgccttaccgtcatatgaatatcctcctt a

2. the building of carrier pDY001

Utilize primer KDC-XbaI (ATCTCTAGATTTAAGAAGGAGATATAATGGCACCTGTTACAATTGAA ) and KDC-NheI-BamHI (TCTGGATCCGCTAGCCTATTTTTTATTTCTTTTAAG TGCCGCTG) PCR amplification AAGTTC The segment of amplification is then inserted into pET28a (+) by the KDC gene from S. cervisiae YPH499 with XbaI and BamHI Obtain carrier pDY01.Utilize primer ALDH-XbaI (ATCTCTAGATTTAAGAAGGAGATATAATGAATTTTCATCATCTGGCTTACTG) and ALDH-NheI-BamHI (TCAGGATCCGCTAGCTCAGGCCTCCAGGCTTATCC) ALDH gene of the PCR amplification from Escherichia coli, using XbaI and BamHI, which is inserted into pET28a (+), obtains carrier pDY02.Utilize primer ARO8-XbaI (AACTCTAGATTTAAGAAGGAGATATAATGATGACTTTACCTGAATCAAAAGACTTT TC) and ARO8-NheI- BamHI (ACAGGATCCGCTAGCCTATTTGGAAATACCAAATTCTTCGTATA A) PCR amplification comes from S. cervisiae The segment of amplification, is then inserted into pET28a (+) with XbaI and BamHI and obtains carrier pDY03 by the KDC gene of YPH499. ALDH expression cassette is obtained using XbaI and XhoI double digestion pDY02, is inserted into the carrier pDY01 with NheI and XhoI double digestion On, obtain carrier pDY04.ARO8 expression cassette is obtained using XbaI and XhoI double digestion pDY03, is inserted into NheI and XhoI On the carrier pDY04 of double digestion, carrier pDY001 is obtained.The KDC gene has nucleotides sequence shown in SEQ ID NO:19 Column coding；The KDC has amino acid sequence shown in SEQ ID NO:20；The ALDH gene has SEQ ID NO:21 institute That shows is nucleotide sequence coded；The ALDH gene has amino acid sequence shown in SEQ ID NO:22；The ARO8 gene With nucleotide sequence coded shown in SEQ ID NO:23；The ARO8 gene has amino acid shown in SEQ ID NO:24 Sequence；

3. the building of carrier pDY002

Release the aroG of feedback inhibition^fbrAnd pheA^fbrGene trust money Wei Zhi Biotechnology Co., Ltd synthesis, with synthesis Gene be template utilize primer aroG^fbr-XbaI(GTATCTAGAAAGAGGAGATATAATGAATTAT ) and aroG CAGAACGACGATTTACGC^fbr-SpeI-BamHI(TATGGATCCACTAGTTTACCCGCGACG CGCTTTTA) PCR amplification aroG^fbrThe segment of amplification is then inserted into pET28a (+) with XbaI and BamHI and obtains carrier by gene pDY05.Using the gene of synthesis as template, primer pheA is utilized^fbrr-XbaI (GTATCTAGAAAGAGGAGATATAATGACATCGGAAAACCCGTTACTG) and pheA^fbr-SpeI-SalI (ATATGTCGACACTAGTTCACAACGTGGTTTTCGCCGGA) PCR amplification carrys out pheA^fbrGene, then by the segment of amplification It is inserted into pET28a (+) with XbaI and SalI and obtains carrier pDY06.It is obtained using XbaI and XhoI double digestion pDY06 pheA^fbrExpression cassette is inserted on the carrier pDY05 with SpeI and XhoI double digestion, obtains carrier pDY07.Using XbaI and XhoI double digestion pDY07 obtains aroG^fbrAnd pheA^fbExpression cassette is inserted into the carrier of XbaI and XhoI double digestion On pBBR1MCS1, carrier pDY002 is obtained.The aroG^fbrGene has nucleotide sequence shown in SEQ ID NO:1；It is described aroG^fbrGene has and amino acid sequence shown in SEQ ID NO:2；The pheA^fbrGene has shown in SEQ ID NO:3 It is nucleotide sequence coded；The pheA^fbrGene has amino acid sequence shown in SEQ ID NO:4；4. by recombinant vector PDY001 and pDY002 is imported in Escherichia coli GDY simultaneously, obtains GDY1 engineering bacteria.The engineering bacteria can realize aroG^fbr、 pheA^fbr, KDC, ALDH and ARO8 gene high expression, effectively can convert phenylacetic acid for monosaccharide.

The building of embodiment 2 produces p-hydroxyphenylaceticacid colibacillus engineering

1. the building of carrier pDY003

Release the tyrA of feedback inhibition^fbrGene trust money Wei Zhi Biotechnology Co., Ltd synthesis, with the gene of the synthesis Primer tyrA is utilized for template^fbrr- XbaI (GTATCTAGAAAGAGGAGATATAATGGTTGCTGAATT GACCGCAT) and tyrA^fbr- SpeI-SalI (ATATGTCGACACTAGTTTACTGGCGATTGTCATTCGCC) PCR amplification carrys out tyrA^fbrGene, The segment of amplification is inserted into pET28a (+) with XbaI and SalI then and obtains carrier pDY08.Utilize the bis- enzymes of XbaI and XhoI It cuts pDY08 and obtains tyrA^fbrExpression cassette is inserted on the carrier pDY05 with SpeI and XhoI double digestion, obtains carrier pDY09.AroG is obtained using XbaI and XhoI double digestion pDY09^fbrAnd tyrA^fbrExpression cassette is inserted into XbaI and XhoI On the carrier pBBR1MCS1 of double digestion, carrier pDY003 is obtained.The tyrA^fbrGene has core shown in SEQ ID NO:5 Nucleotide sequence coding；The tyrA^fbrGene has amino acid sequence shown in SEQ ID NO:6；

2. recombinant vector pDY001 and pDY003 is imported in Escherichia coli GDY simultaneously, GDY2 engineering bacteria is obtained.The engineering Bacterium can realize aroG^fbr、tyrA^fbr, KDC, ALDH and ARO8 gene high expression, effectively can convert para hydroxybenzene for monosaccharide Acetic acid.

The building of embodiment 3 produces heteroauxin colibacillus engineering

1. the building of carrier pDY004

Release the TrpE of feedback inhibition^fbrAnd SerA^fbrThe synthesis of gene trust money Wei Zhi Biotechnology Co., Ltd.With synthesis Gene be template, utilize primer TrpE^fbr-XbaI (GTATCTAGAAAGAGGAGATATAATGCAAACACAAAAACCGACTCTC) and TrpE^fbr-SpeI-BamHI (TATGGATCCACTAGTTCAGAAAGTCTCCTGTGCATGATG) PCR amplification TrpE^fbrGene, then by the segment of amplification with XbaI and BamHI, which is inserted into pET28a (+), obtains carrier pDY10.Using the gene of synthesis as template, primer SerA is utilized^fbr- XbaI (GTATCTAGAAAGAGGAGATATAATGACATCG GAAAACCCGTTACTG) and SerA^fbr-SpeI-SalI (ATATGTCGACACTAGTTCACAACGTGGTTTT CGCCGGA) PCR amplification carrys out SerA^fbrGene, then by the segment of amplification It is inserted into pET28a (+) with XbaI and SalI and obtains carrier pDY11.It is obtained using XbaI and XhoI double digestion pDY10 TrpE^fbrExpression cassette is inserted on the carrier pDY01 with SpeI and XhoI double digestion, obtains carrier pDY12.Using XbaI and XhoI double digestion pDY10 obtains TrpE^fbrExpression cassette is inserted on the carrier pDY12 with SpeI and XhoI double digestion, obtains Carrier pDY13.SerA is obtained using XbaI and XhoI double digestion pDY13^fbrAnd TrpE^fbrExpression cassette, be inserted into XbaI and On the carrier pBBR1MCS1 of XhoI double digestion, carrier pDY004 is obtained.The TrpE^fbrGene has shown in SEQ ID NO:7 It is nucleotide sequence coded；The TrpE^fbrGene has amino acid sequence shown in SEQ ID NO:8；serA^fbrIt is described serA^fbrGene has nucleotide sequence coded shown in SEQ ID NO:9；The serA^fbrGene has SEQ ID NO:10 Shown in amino acid sequence；

2. the building of carrier pDY005

Using Escherichia coli MG1655 genome as template, primer TrpABCD-XbaI is utilized (CCATCTAGATTTAAGAGGAGATATAATGGCTGACATTCTGCTGC TCGATA) and TrpABCD-XhoI (GATTCTCGAGTTAACTGCGCGTCGCCGCT) PCR amplification TrpABCD gene (includes trpA, trpB, trpC and trpD tetra- A gene), the segment of amplification is inserted into pDY001 with XbaI and XhoI then, obtains carrier pDY005.The trpA tool Have nucleotide sequence coded shown in SEQ ID NO:11；TrpA gene has amino acid sequence shown in SEQ ID NO:12； The trpB has nucleotide sequence coded shown in SEQ ID NO:13；The trpB gene has shown in SEQ ID NO:14 Amino acid sequence；The trpC has nucleotide sequence coded shown in SEQ ID NO:15；The trpC gene has SEQ Amino acid sequence shown in ID NO:16；The trpD has nucleotide sequence coded described shown in SEQ ID NO:17；Institute TrpD gene is stated with amino acid sequence shown in SEQ ID NO:18；

3. recombinant vector pDY004 and pDY005 is imported in Escherichia coli GDY simultaneously, GDY3 engineering bacteria is obtained.The engineering Bacterium can realize aroG^fbr、TrpE^fbr、SerA^fbr, TrpABCD, KDC, ALDH and ARO8 gene high expression, can effectively will be single Sugar is converted into heteroauxin.

The building of embodiment 4 produces auximone Pichia yeast engineering GDY4

Using Escherichia coli MG1655 genome as template, primer ALDH-BstBI (AGCATTCGAAAT is utilized ) and ALDH-XhoI (GAACCTCGAGTCAGGCCTCCAGGCTTAT CCAGAT) GAATTTTCATCATCTGGCTTACTGGC The segment of amplification, is then inserted into pGAPZa with BstBI and XhoI I and obtains carrier pDY006 by PCR amplification ALDHr gene. Using saccharomyces cerevisiae YPH499 genome as template, primer KDC-BstBI is utilized (AGCATTCGAAATGGCACCTGTTACAATTGAAAAGTTC)/KDC-XhoI(CTTCCTC GAGCTATTTTTTATTTCTTTTAAGTGCCGCTG), primer ARO8-BstBI (AGCATTCGAAATGACTT TACCTGAATCAAAAGACTTTTC)/ARO8-XhoI(CTTCCTCGAGCTATTTGGAAATACCAAATTCTTC GTATAA) The segment of amplification, is then inserted respectively into pGAPZa with BstBI and XhoI, obtains carrier by PCR amplification KDC and ARO8 gene PDY14 and pDY15.By pDY14 and pDY15 respectively with BamHI and BglII double digestion, KDC gene expression frame is obtained (comprising GAP Promoter, KDC gene and AOX1TT) and ARO8 gene expression frame (including GAP promoter, ARO8 gene and AOX1TT).By KDC Gene expression frame is recombinated to the pAO815 carrier through BamHI digestion, obtains carrier pDY007.The recombination of ARO8 gene expression frame is arrived PPIC3.5K carrier through BamHI digestion, obtains carrier pDY008.Carrier pDY006, pDY007 and pDY008 are distinguished linear It after change, successively imports in Pichia pastoris GS115, realizes the constitutive expression of ARO8, ALDH and KDC gene, engineering bacteria name For GDY4.Expression ARO8 turns to phenylpyruvic acid, ethyl-para-hydroxyphenyl ketone and Yin for being catalyzed phenylalanine, tyrosine and tryptophan transfer Diindyl pyruvic acid.Expressing K DC for be catalyzed phenylpyruvic acid, Single-chip microcomputer and indolepyruvic acid be separately converted to phenylacetaldehyde, Para hydroxybenzene acetaldehyde and indoleacetaldehyde.Expression ALDH is converted respectively for being catalyzed phenylacetaldehyde, para hydroxybenzene acetaldehyde and indoleacetaldehyde For phenylacetic acid, p-hydroxyphenylaceticacid and heteroauxin.There is the KDC gene nucleotide sequence shown in SEQ ID NO:19 to compile Code；The KDC has amino acid sequence shown in SEQ ID NO:20；The ALDH gene has shown in SEQ ID NO:21 It is nucleotide sequence coded；The ALDH gene has amino acid sequence shown in SEQ ID NO:22；The ARO8 gene has It is nucleotide sequence coded shown in SEQ ID NO:23；The ARO8 gene has amino acid sequence shown in SEQ ID NO:24 Column；

The fermenting experiment of 5 colibacillus engineering of embodiment production auximone

1, colibacillus engineering GDY1, GDY2 and GDY3 that will be incubated overnight in 37 degree of LB, is inoculated into 50ml M9 respectively In fermentation medium (including 2% glucose), 30 degree of shake flask fermentations are carried out, to thallus OD₆₀₀It is 0.8 or so, 0.1mM is added The inducing expression of IPTG progress target gene.

2, fermented and cultured collects thallus, ultrasonic wave broken cell after 24 hours, and product is extracted with ethyl acetate in benefit.

3, solvent ethyl acetate is spin-dried for using Rotary Evaporators, then with a certain amount of ethyl acetate lysate.

4, GC-MS (gas chromatograph-mass spectrometer (GC-MS)) test sample.

5, experimental result: colibacillus engineering is using the GC-MS testing result of glucose synthetic auxin as schemed 1, shown in Fig. 2 and Fig. 3.Fig. 1 is colibacillus engineering GDY1 tunning GC-MS testing result, shows that bacterial strain GDY1 can have Effect utilizes glucose synthetic auxin phenylacetic acid, and shaking table yield is 1.67g/L.Fig. 2 is colibacillus engineering GDY2 hair Ferment product GC-MS testing result shows that bacterial strain GDY2 can efficiently use glucose synthetic auxin p-hydroxyphenylaceticacid, shakes Bed yield is 1.07g/L.Fig. 3 is colibacillus engineering GDY3 tunning GC-MS testing result, shows that bacterial strain GDY3 can have For effect using glucose synthetic auxin to heteroauxin, shaking table yield is 1.16g/L.

The fermenting experiment of 6 Pichia yeast engineering of embodiment production auximone

6, the Pichia yeast engineering GDY4 that will be incubated overnight in 30 degree of YPD is inoculated into 50ml YPD fermentation medium (including 2% glucose), carries out 30 degree of shake flask fermentations.

7, fermented and cultured collects thallus, ultrasonic wave broken cell after 28 hours, and product is extracted with ethyl acetate in benefit.

8, solvent ethyl acetate is spin-dried for using Rotary Evaporators, then with a certain amount of ethyl acetate lysate.

9, GC-MS (gas chromatograph-mass spectrometer (GC-MS)) test sample.

10, experimental result: Pichia yeast engineering is using the GC-MS testing result of glucose synthetic auxin as schemed Shown in 4.The result shows that bacterial strain GDY4 can efficiently use glucose synthetic auxin phenylacetic acid, p-hydroxyphenylaceticacid and indoles Acetic acid, yield are respectively 108mg/L, 36mg/L and 33mg/L.

The above is only the preferred embodiments of the invention, it is noted that for those skilled in the art, Under the premise of not departing from this clearly demarcated principle, it can also make and be suitably modified, these improvement also should be regarded as protection model of the invention It encloses.

Sequence table

<110>south jiangxi normal university

<120>a kind of method of Microbe synthesis auximone

<160> 55

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1053

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 1

atgaattatc agaacgacga tttacgcatc aaagaaatca aagagttact tcctcctgtc 60

gcattgctgg aaaaattccc cgctactgaa aatgccgcga atacggttgc ccatgcccga 120

aaagcgatcc ataagatcct gaaaggtaat gatgatcgcc tgttggttgt gattggccca 180

tgctcaattc atgatcctgt cgcggcaaaa gagtatgcca ctcgcttgct ggcgctgcgt 240

gaagagctga aagatgagct ggaaatcgta atgcgcgtct attttgaaaa gccgcgtacc 300

acggtgggct ggaaagggct gattaacgat ccgcatatgg ataatagctt ccagatcaac 360

gacggtctgc gtatagcccg taaattgctg cttgatatta acgacagcgg tctgccagcg 420

gcaggtgagt ttctcaatat gatcacccca caatatctcg ctgacctgat gagctggggc 480

gcaattggcg cacgtaccac cgaatcgcag gtgcaccgcg aactggcatc agggctttct 540

tgtccggtcg gcttcaaaaa tggcaccgac ggtacgatta aagtggctat cgatgccatt 600

aatgccgccg gtgcgccgca ctgcttcctg tccgtaacga aatgggggca ttcggcgatt 660

gtgaatacca gcggtaacgg cgattgccat atcattctgc gcggcggtaa agagcctaac 720

tacagcgcga agcacgttgc tgaagtgaaa gaagggctga acaaagcagg cctgccagca 780

caggtgatga tcgatttcag ccatgctaac tcgtccaaac aattcaaaaa gcagatggat 840

gtttgtgctg acgtttgcca gcagattgcc ggtggcgaaa aggccattat tggcgtgatg 900

gtggaaagcc atctggtgga aggcaatcag agcctcgaga gcggggagcc gctggcctac 960

ggtaagagca tcaccgatgc ctgcatcggc tgggaagata ccgatgctct gttacgtcaa 1020

ctggcgaatg cagtaaaagc gcgtcgcggg taa 1053

<210> 3

<211> 350

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 3

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

1 5 10 15

Leu Pro Pro Val Ala Leu Leu Glu Lys Phe Pro Ala Thr Glu Asn Ala

20 25 30

Ala Asn Thr Val Ala His Ala Arg Lys Ala Ile His Lys Ile Leu Lys

35 40 45

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

50 55 60

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

65 70 75 80

Glu Glu Leu Lys Asp Glu Leu Glu Ile Val Met Arg Val Tyr Phe Glu

85 90 95

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

100 105 110

Met Asp Asn Ser Phe Gln Ile Asn Asp Gly Leu Arg Ile Ala Arg Lys

115 120 125

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

130 135 140

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

145 150 155 160

Ala Ile Gly Ala Arg Thr Thr Glu Ser Gln Val His Arg Glu Leu Ala

165 170 175

Ser Gly Leu Ser Cys Pro Val Gly Phe Lys Asn Gly Thr Asp Gly Thr

180 185 190

Ile Lys Val Ala Ile Asp Ala Ile Asn Ala Ala Gly Ala Pro His Cys

195 200 205

Phe Leu Ser Val Thr Lys Trp Gly His Ser Ala Ile Val Asn Thr Ser

210 215 220

Gly Asn Gly Asp Cys His Ile Ile Leu Arg Gly Gly Lys Glu Pro Asn

225 230 235 240

Tyr Ser Ala Lys His Val Ala Glu Val Lys Glu Gly Leu Asn Lys Ala

245 250 255

Gly Leu Pro Ala Gln Val Met Ile Asp Phe Ser His Ala Asn Ser Ser

260 265 270

Lys Gln Phe Lys Lys Gln Met Asp Val Cys Ala Asp Val Cys Gln Gln

275 280 285

Ile Ala Gly Gly Glu Lys Ala Ile Ile Gly Val Met Val Glu Ser His

290 295 300

Leu Val Glu Gly Asn Gln Ser Leu Glu Ser Gly Glu Pro Leu Ala Tyr

305 310 315 320

Gly Lys Ser Ile Thr Asp Ala Cys Ile Gly Trp Glu Asp Thr Asp Ala

325 330 335

Leu Leu Arg Gln Leu Ala Asn Ala Val Lys Ala Arg Arg Gly

340 345 350

<210> 3

<211> 903

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 3

atgacatcgg aaaacccgtt actggcgctg cgagagaaaa tcagcgcgct ggatgaaaaa 60

ttattagcgt tactggcaga acggcgcgaa ctggccgtcg aggtgggaaa agccaaactg 120

ctctcgcatc gcccggtacg tgatattgat cgtgaacgcg atttgctgga aagattaatt 180

acgctcggta aagcgcacca tctggacgcc cattacatta ctcgcctgtt ccagctcatc 240

attgaagatt ccgtattaac tcagcaggct ttgctccaac aacatctcaa taaaattaat 300

ccgcactcag cacgcatcgc ttttctcggc cccaaaggtt cttattccca tcttgcggcg 360

cgccagtatg ctgcccgtca ctttgagcaa ttcattgaaa gtggctgcgc caaatttgcc 420

gatattttta atcaggtgga aaccggccag gccgactatg ccgtcgtacc gattgaaaat 480

accagctccg gtgccataaa cgacgtttac gatctgctgc aacataccag cttgtcgatt 540

gttggcgaga tgacgttaac tatcgaccat tgtttgttgg tctccggcac tactgattta 600

tccaccatca atacggtcta cagccatccg cagccattcc agcaatgcag caaattcctt 660

aatcgttatc cgcactggaa gattgaatat accgaaagta cgtctgcggc aatggaaaag 720

gttgcacagg caaaatcacc gcatgttgct gcgttgggaa gcgaagctgg cggcactttg 780

tacggtttgc aggtactgga gcgtattgaa gcaaatcagc gacaaaactt cacccgattt 840

gtggtgttgg cgcgtaaagc cattaacgtg tctgatcagg ttccggcgaa aaccacgttg 900

tga 903

<210> 4

<211> 300

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 4

Met Thr Ser Glu Asn Pro Leu Leu Ala Leu Arg Glu Lys Ile Ser Ala

1 5 10 15

Leu Asp Glu Lys Leu Leu Ala Leu Leu Ala Glu Arg Arg Glu Leu Ala

20 25 30

Val Glu Val Gly Lys Ala Lys Leu Leu Ser His Arg Pro Val Arg Asp

35 40 45

Ile Asp Arg Glu Arg Asp Leu Leu Glu Arg Leu Ile Thr Leu Gly Lys

50 55 60

Ala His His Leu Asp Ala His Tyr Ile Thr Arg Leu Phe Gln Leu Ile

65 70 75 80

Ile Glu Asp Ser Val Leu Thr Gln Gln Ala Leu Leu Gln Gln His Leu

85 90 95

Asn Lys Ile Asn Pro His Ser Ala Arg Ile Ala Phe Leu Gly Pro Lys

100 105 110

Gly Ser Tyr Ser His Leu Ala Ala Arg Gln Tyr Ala Ala Arg His Phe

115 120 125

Glu Gln Phe Ile Glu Ser Gly Cys Ala Lys Phe Ala Asp Ile Phe Asn

130 135 140

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

145 150 155 160

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

165 170 175

Ser Leu Ser Ile Val Gly Glu Met Thr Leu Thr Ile Asp His Cys Leu

180 185 190

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

195 200 205

His Pro Gln Pro Phe Gln Gln Cys Ser Lys Phe Leu Asn Arg Tyr Pro

210 215 220

His Trp Lys Ile Glu Tyr Thr Glu Ser Thr Ser Ala Ala Met Glu Lys

225 230 235 240

Val Ala Gln Ala Lys Ser Pro His Val Ala Ala Leu Gly Ser Glu Ala

245 250 255

Gly Gly Thr Leu Tyr Gly Leu Gln Val Leu Glu Arg Ile Glu Ala Asn

260 265 270

Gln Arg Gln Asn Phe Thr Arg Phe Val Val Leu Ala Arg Lys Ala Ile

275 280 285

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

290 295 300

<210> 5

<211> 1122

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 5

atggttgctg aattgaccgc attacgcgat caaattgatg aagtcgataa agcgctgctg 60

aatttattag cgaagcgtct ggaactggtt gctgaagtgg gcgaggtgaa aagccgcttt 120

ggactgccta tttatgttcc ggagcgcgag gcatctattt tggcctcgcg gcgcgcagag 180

gcggaagctc tgggtgtacc gccagatctg attgaggatg ttttgcgtcg ggtgatgcgt 240

gaatcttact ccagtgaaaa cgacaaagga tttaaaacac tttgtccgtc actgcgtccg 300

gtggttatcg tcggcggtgg cggtcagatg ggacgcctgt tcgagaagat gctgacacta 360

tcgggttatc aggtgcggat tctggagcaa catgactggg atcgagcggc tgatattgtt 420

gccgatgccg gaatggtgat tgttagtgtg ccaatccacg ttactgagca agttatcggc 480

aaattaccgc ctttaccgaa agattgtatt ctggttgatc tggcatcagt gaaaaatgga 540

ccattacagg ccatgctggc ggcgcacgat ggcccggtac tggggttaca cccgatgttc 600

ggcccggaca gcggtagcct ggcaaagcaa gttgtggtct ggtgtgatgg acgtaagccg 660

gaagcatacc aatggtttct ggagcaaatt caggtctggg gcgctcggct gcatcgtatt 720

agcgctgtcg agcacgatca gaatatggcg tttattcagg ctctgcgcca ctttgctact 780

tttgcttatg ggctgcatct ggcagaagaa aatgttcagc ttgagcaact tctggcgctc 840

tcttcgccga tttaccgcct tgagctggcg atggtcgggc gactgtttgc tcaggatccg 900

cagctttatg ccgacattat tatgtcgtca gagcgtaatc tggcgttaat caaacgttac 960

tataagcgtt tcggcgaggc gattgagttg ctggagcagg gcgataagca ggcgtttatt 1020

gacagtttcc gcaaggtgga gcactggttc ggcgattacg ttcagcgttt tcagagtgaa 1080

agccgcgtgt tattgcgtca ggcgaatgac aatcgccagt aa 1122

<210> 6

<211> 373

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 6

Met Val Ala Glu Leu Thr Ala Leu Arg Asp Gln Ile Asp Glu Val Asp

1 5 10 15

Lys Ala Leu Leu Asn Leu Leu Ala Lys Arg Leu Glu Leu Val Ala Glu

20 25 30

Val Gly Glu Val Lys Ser Arg Phe Gly Leu Pro Ile Tyr Val Pro Glu

35 40 45

Arg Glu Ala Ser Ile Leu Ala Ser Arg Arg Ala Glu Ala Glu Ala Leu

50 55 60

Gly Val Pro Pro Asp Leu Ile Glu Asp Val Leu Arg Arg Val Met Arg

65 70 75 80

Glu Ser Tyr Ser Ser Glu Asn Asp Lys Gly Phe Lys Thr Leu Cys Pro

85 90 95

Ser Leu Arg Pro Val Val Ile Val Gly Gly Gly Gly Gln Met Gly Arg

100 105 110

Leu Phe Glu Lys Met Leu Thr Leu Ser Gly Tyr Gln Val Arg Ile Leu

115 120 125

Glu Gln His Asp Trp Asp Arg Ala Ala Asp Ile Val Ala Asp Ala Gly

130 135 140

Met Val Ile Val Ser Val Pro Ile His Val Thr Glu Gln Val Ile Gly

145 150 155 160

Lys Leu Pro Pro Leu Pro Lys Asp Cys Ile Leu Val Asp Leu Ala Ser

165 170 175

Val Lys Asn Gly Pro Leu Gln Ala Met Leu Ala Ala His Asp Gly Pro

180 185 190

Val Leu Gly Leu His Pro Met Phe Gly Pro Asp Ser Gly Ser Leu Ala

195 200 205

Lys Gln Val Val Val Trp Cys Asp Gly Arg Lys Pro Glu Ala Tyr Gln

210 215 220

Trp Phe Leu Glu Gln Ile Gln Val Trp Gly Ala Arg Leu His Arg Ile

225 230 235 240

Ser Ala Val Glu His Asp Gln Asn Met Ala Phe Ile Gln Ala Leu Arg

245 250 255

His Phe Ala Thr Phe Ala Tyr Gly Leu His Leu Ala Glu Glu Asn Val

260 265 270

Gln Leu Glu Gln Leu Leu Ala Leu Ser Ser Pro Ile Tyr Arg Leu Glu

275 280 285

Leu Ala Met Val Gly Arg Leu Phe Ala Gln Asp Pro Gln Leu Tyr Ala

290 295 300

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

305 310 315 320

Tyr Lys Arg Phe Gly Glu Ala Ile Glu Leu Leu Glu Gln Gly Asp Lys

325 330 335

Gln Ala Phe Ile Asp Ser Phe Arg Lys Val Glu His Trp Phe Gly Asp

340 345 350

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

355 360 365

Asn Asp Asn Arg Gln

370

<210> 7

<211> 1563

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 7

atgcaaacac aaaaaccgac tctcgaacag ctaacctgcg aaggcgctta tcgcgacaat 60

cccaccgcgc tttttcacca gttgtgtggg gatcgtccgg caacgctgct gctggaattt 120

gcagatatcg acagcaaaga tgatttaaaa agcctgctgc tggtagacag tgcgctgcgc 180

attacagctt taggtgacac tgtcacaatc caggcacttt ccggcaacgg cgaagccctg 240

ctggcactac tggataacgc cctgcctgcg ggtgtggaaa gtgaacaatc accaaactgc 300

cgtgtgctgc gcttcccccc tgtcagtcca ctgctggatg aagacgcccg cttatgctcc 360

ctttcggttt ttgacgcttt ccgtttattg cagaatctgt tgaatgtacc gaaggaagaa 420

cgagaagcca tgttcttcgg cggcctgttc tcttatgacc ttgtggcggg atttgaagat 480

ttaccgcaac tgtcagcgga aaataactgc cctgatttct gtttttatct cgctgaaacg 540

ctgatggtga ttgaccatca gaaaaaaagc acccgtattc aggccagcct gtttgctccg 600

aatgaagaag aaaaacaacg tctcactgct cgcctgaacg aactacgtca gcaactgacc 660

gaagccgcgc cgccgctgcc agtggtttcc gtgccgcata tgcgttgtga atgtaatcag 720

agcgatgaag agttcggtgg cgtagtgcgt ttgttgcaaa aagcgattcg cgctggagaa 780

attttccagg tggtgccatc tcgccgtttc tctctgccct gcccgtcacc gctggcggcc 840

tattacgtgc tgaaaaagag taatcccagc ccgtacatgt tttttatgca ggataatgat 900

ttcaccctat ttggcgcgtc gccggaaagc tcgctcaagt atgatgccac cagccgccag 960

attgagatct acccgattgc cggaacacgc ccacgcggtc gtcgcgccga tggttcactg 1020

gacagagatc tcgacagccg tattgaactg gaaatgcgta ccgatcataa agagctgtct 1080

gaacatctga tgctggttga tctcgcccgt aatgatctgg cacgcatttg cacccccggc 1140

agccgctacg tcgccgatct caccaaagtt gaccgttatt cctatgtgat gcacctcgtc 1200

tctcgcgtag tcggcgaact gcgtcacgat cttgacgccc tgcacgctta tcgcgcctgt 1260

atgaatatgg ggacgttaag cggtgcgccg aaagtacgcg ctatgcagtt aattgccgag 1320

gcggaaggtc gtcgccgcgg cagctacggc ggcgcggtag gttatttcac cgcgcatggc 1380

gatctcgaca cctgcattgt gatccgctcg gcgctggtgg aaaacggtat cgccaccgtg 1440

caagcgggtg ctggtgtagt ccttgattct gttccgcagt cggaagccga cgaaacccgt 1500

aacaaagccc gcgctgtact gcgcgctatt gccaccgcgc atcatgcaca ggagactttc 1560

tga 1563

<210> 8

<211> 520

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 8

Met Gln Thr Gln Lys Pro Thr Leu Glu Gln Leu Thr Cys Glu Gly Ala

1 5 10 15

Tyr Arg Asp Asn Pro Thr Ala Leu Phe His Gln Leu Cys Gly Asp Arg

20 25 30

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

35 40 45

Leu Lys Ser Leu Leu Leu Val Asp Ser Ala Leu Arg Ile Thr Ala Leu

50 55 60

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

65 70 75 80

Leu Ala Leu Leu Asp Asn Ala Leu Pro Ala Gly Val Glu Ser Glu Gln

85 90 95

Ser Pro Asn Cys Arg Val Leu Arg Phe Pro Pro Val Ser Pro Leu Leu

100 105 110

Asp Glu Asp Ala Arg Leu Cys Ser Leu Ser Val Phe Asp Ala Phe Arg

115 120 125

Leu Leu Gln Asn Leu Leu Asn Val Pro Lys Glu Glu Arg Glu Ala Met

130 135 140

Phe Phe Gly Gly Leu Phe Ser Tyr Asp Leu Val Ala Gly Phe Glu Asp

145 150 155 160

Leu Pro Gln Leu Ser Ala Glu Asn Asn Cys Pro Asp Phe Cys Phe Tyr

165 170 175

Leu Ala Glu Thr Leu Met Val Ile Asp His Gln Lys Lys Ser Thr Arg

180 185 190

Ile Gln Ala Ser Leu Phe Ala Pro Asn Glu Glu Glu Lys Gln Arg Leu

195 200 205

Thr Ala Arg Leu Asn Glu Leu Arg Gln Gln Leu Thr Glu Ala Ala Pro

210 215 220

Pro Leu Pro Val Val Ser Val Pro His Met Arg Cys Glu Cys Asn Gln

225 230 235 240

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

245 250 255

Arg Ala Gly Glu Ile Phe Gln Val Val Pro Ser Arg Arg Phe Ser Leu

260 265 270

Pro Cys Pro Ser Pro Leu Ala Ala Tyr Tyr Val Leu Lys Lys Ser Asn

275 280 285

Pro Ser Pro Tyr Met Phe Phe Met Gln Asp Asn Asp Phe Thr Leu Phe

290 295 300

Gly Ala Ser Pro Glu Ser Ser Leu Lys Tyr Asp Ala Thr Ser Arg Gln

305 310 315 320

Ile Glu Ile Tyr Pro Ile Ala Gly Thr Arg Pro Arg Gly Arg Arg Ala

325 330 335

Asp Gly Ser Leu Asp Arg Asp Leu Asp Ser Arg Ile Glu Leu Glu Met

340 345 350

Arg Thr Asp His Lys Glu Leu Ser Glu His Leu Met Leu Val Asp Leu

355 360 365

Ala Arg Asn Asp Leu Ala Arg Ile Cys Thr Pro Gly Ser Arg Tyr Val

370 375 380

Ala Asp Leu Thr Lys Val Asp Arg Tyr Ser Tyr Val Met His Leu Val

385 390 395 400

Ser Arg Val Val Gly Glu Leu Arg His Asp Leu Asp Ala Leu His Ala

405 410 415

Tyr Arg Ala Cys Met Asn Met Gly Thr Leu Ser Gly Ala Pro Lys Val

420 425 430

Arg Ala Met Gln Leu Ile Ala Glu Ala Glu Gly Arg Arg Arg Gly Ser

435 440 445

Tyr Gly Gly Ala Val Gly Tyr Phe Thr Ala His Gly Asp Leu Asp Thr

450 455 460

Cys Ile Val Ile Arg Ser Ala Leu Val Glu Asn Gly Ile Ala Thr Val

465 470 475 480

Gln Ala Gly Ala Gly Val Val Leu Asp Ser Val Pro Gln Ser Glu Ala

485 490 495

Asp Glu Thr Arg Asn Lys Ala Arg Ala Val Leu Arg Ala Ile Ala Thr

500 505 510

Ala His His Ala Gln Glu Thr Phe

515 520

<210> 9

<211> 1233

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 9

atggcaaagg tatcgctgga gaaagacaag attaagtttc tgctggtaga aggcgtgcac 60

caaaaggcgc tggaaagcct tcgtgcagct ggttacacca acatcgaatt tcacaaaggc 120

gcgctggatg atgaacaatt aaaagaatcc atccgcgatg cccacttcat cggcctgcga 180

tcccgtaccc atctgactga agacgtgatc aacgccgcag aaaaactggt cgctattggc 240

tgtttctgta tcggaacaaa ccaggttgat ctggatgcgg cggcaaagcg cgggatcccg 300

gtatttaacg caccgttctc aaatacgcgc tctgttgcgg agctggtgat tggcgaactg 360

ctgctgctat tgcgcggcgt gccggaagcc aatgctaaag cgcaccgtgg cgtgtggaac 420

aaactggcgg cgggttcttt tgaagcgcgc ggcaaaaagc tgggtatcat cggctacggt 480

catattggta cgcaattggg cattctggct gaatcgctgg gaatgtatgt ttacttttat 540

gatattgaaa ataaactgcc gctgggcaac gccactcagg tacagcatct ttctgacctg 600

ctgaatatga gcgatgtggt gagtctgcat gtaccagaga atccgtccac caaaaatatg 660

atgggcgcga aagaaatttc actaatgaag cccggctcgc tgctgattaa tgcttcgcgc 720

ggtactgtgg tggatattcc ggcgctgtgt gatgcgctgg cgagcaaaca tctggcgggg 780

gcggcaatcg acgtattccc gacggaaccg gcgaccaata gcgatccatt tacctctccg 840

ctgtgtgaat tcgacaacgt ccttctgacg ccacacattg gcggttcgac tcaggaagcg 900

caggagaata tcggcctgga agttgcgggt aaattgatca agtattctga caatggctca 960

acgctctctg cggtgaactt cccggaagtc tcgctgccac tgcacggtgg gcgtcgtctg 1020

atgcacatcg cagaaaaccg tccgggcgtg ctaactgcgc tgaacaaaat cttcgccgag 1080

cagggcgtcg caatcgccgc gcaatatctg caaacttccg cccagatggg ttatgtggtt 1140

attgatattg aagccgacga agacgttgcc gaaaaagcgc tgcaggcaat gaaagctatt 1200

ccgggtacca ttcgcgcccg tctgctgtac taa 1233

<210> 10

<211> 410

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 10

Met Ala Lys Val Ser Leu Glu Lys Asp Lys Ile Lys Phe Leu Leu Val

1 5 10 15

Glu Gly Val His Gln Lys Ala Leu Glu Ser Leu Arg Ala Ala Gly Tyr

20 25 30

Thr Asn Ile Glu Phe His Lys Gly Ala Leu Asp Asp Glu Gln Leu Lys

35 40 45

Glu Ser Ile Arg Asp Ala His Phe Ile Gly Leu Arg Ser Arg Thr His

50 55 60

Leu Thr Glu Asp Val Ile Asn Ala Ala Glu Lys Leu Val Ala Ile Gly

65 70 75 80

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

85 90 95

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

100 105 110

Ala Glu Leu Val Ile Gly Glu Leu Leu Leu Leu Leu Arg Gly Val Pro

115 120 125

Glu Ala Asn Ala Lys Ala His Arg Gly Val Trp Asn Lys Leu Ala Ala

130 135 140

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

145 150 155 160

His Ile Gly Thr Gln Leu Gly Ile Leu Ala Glu Ser Leu Gly Met Tyr

165 170 175

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

180 185 190

Gln Val Gln His Leu Ser Asp Leu Leu Asn Met Ser Asp Val Val Ser

195 200 205

Leu His Val Pro Glu Asn Pro Ser Thr Lys Asn Met Met Gly Ala Lys

210 215 220

Glu Ile Ser Leu Met Lys Pro Gly Ser Leu Leu Ile Asn Ala Ser Arg

225 230 235 240

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

245 250 255

His Leu Ala Gly Ala Ala Ile Asp Val Phe Pro Thr Glu Pro Ala Thr

260 265 270

Asn Ser Asp Pro Phe Thr Ser Pro Leu Cys Glu Phe Asp Asn Val Leu

275 280 285

Leu Thr Pro His Ile Gly Gly Ser Thr Gln Glu Ala Gln Glu Asn Ile

290 295 300

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

305 310 315 320

Thr Leu Ser Ala Val Asn Phe Pro Glu Val Ser Leu Pro Leu His Gly

325 330 335

Gly Arg Arg Leu Met His Ile Ala Glu Asn Arg Pro Gly Val Leu Thr

340 345 350

Ala Leu Asn Lys Ile Phe Ala Glu Gln Gly Val Ala Ile Ala Ala Gln

355 360 365

Tyr Leu Gln Thr Ser Ala Gln Met Gly Tyr Val Val Ile Asp Ile Glu

370 375 380

Ala Asp Glu Asp Val Ala Glu Lys Ala Leu Gln Ala Met Lys Ala Ile

385 390 395 400

Pro Gly Thr Ile Arg Ala Arg Leu Leu Tyr

405 410

<210> 11

<211> 807

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 11

atggaacgct acgaatctct gtttgcccag ttgaaggagc gcaaagaagg cgcattcgtt 60

cctttcgtca cgctcggtga tccgggcatt gagcagtcat tgaaaattat cgatacgcta 120

attgaagccg gtgctgacgc gctggagtta ggtatcccct tctccgaccc actggcggat 180

ggcccgacga ttcaaaacgc cactctgcgc gcctttgcgg caggtgtgac tccggcacaa 240

tgttttgaaa tgctggcact gattcgccag aaacacccga ccattcccat tggcctgttg 300

atgtatgcca atctggtgtt taacaaaggc attgatgagt tttatgccca gtgcgaaaaa 360

gtcggcgtcg attcggtgct ggttgccgat gtgccagttg aagagtccgc gcccttccgc 420

caggccgcgt tgcgtcataa tgtcgcacct atcttcatct gcccgccaaa tgccgatgac 480

gacctgctgc gccagatagc ctcttacggt cgtggttaca cctatttgct gtcacgagca 540

ggcgtgaccg gcgcagaaaa ccgcgccgcg ttacccctca atcatctggt tgcgaagctg 600

aaagagtaca acgctgcacc tccattgcag ggatttggta tttccgcccc ggatcaggta 660

aaagcagcga ttgatgcagg agctgcgggc gcgatttctg gttcggccat tgttaaaatc 720

atcgagcaac atattaatga gccagagaaa atgctggcgg cactgaaagt ttttgtacaa 780

ccgatgaaag cggcgacgcg cagttaa 807

<210> 12

<211> 268

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 12

Met Glu Arg Tyr Glu Ser Leu Phe Ala Gln Leu Lys Glu Arg Lys Glu

1 5 10 15

Gly Ala Phe Val Pro Phe Val Thr Leu Gly Asp Pro Gly Ile Glu Gln

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Cys Phe Glu Met Leu Ala Leu Ile Arg Gln Lys His Pro Thr Ile Pro

85 90 95

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

100 105 110

Glu Phe Tyr Ala Gln Cys Glu Lys Val Gly Val Asp Ser Val Leu Val

115 120 125

Ala Asp Val Pro Val Glu Glu Ser Ala Pro Phe Arg Gln Ala Ala Leu

130 135 140

Arg His Asn Val Ala Pro Ile Phe Ile Cys Pro Pro Asn Ala Asp Asp

145 150 155 160

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

165 170 175

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

180 185 190

Leu Asn His Leu Val Ala Lys Leu Lys Glu Tyr Asn Ala Ala Pro Pro

195 200 205

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

210 215 220

Asp Ala Gly Ala Ala Gly Ala Ile Ser Gly Ser Ala Ile Val Lys Ile

225 230 235 240

Ile Glu Gln His Ile Asn Glu Pro Glu Lys Met Leu Ala Ala Leu Lys

245 250 255

Val Phe Val Gln Pro Met Lys Ala Ala Thr Arg Ser

260 265

<210> 13

<211> 1194

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 13

atgacaacat tacttaaccc ctattttggt gagtttggcg gcatgtacgt gccacaaatc 60

ctgatgcctg ctctgcgcca gctggaagaa gcttttgtca gtgcgcaaaa agatcctgaa 120

tttcaggctc agttcaacga cctgctgaaa aactatgccg ggcgtccaac cgcgctgacc 180

aaatgccaga acattacagc cgggacgaac accacgctgt atctcaagcg tgaagatttg 240

ctgcacggcg gcgcgcataa aactaaccag gtgctggggc aggcgttgct ggcgaagcgg 300

atgggtaaaa ccgaaatcat cgccgaaacc ggtgccggtc agcatggcgt ggcgtcggcc 360

cttgccagcg ccctgctcgg cctgaaatgc cgtatttata tgggtgccaa agacgttgaa 420

cgccagtcgc ctaacgtttt tcgtatgcgc ttaatgggtg cggaagtgat cccggtgcat 480

agcggttccg cgacgctgaa agatgcctgt aacgaggcgc tgcgcgactg gtccggtagt 540

tacgaaaccg cgcactatat gctgggcacc gcagctggcc cgcatcctta tccgaccatt 600

gtgcgtgagt ttcagcggat gattggcgaa gaaaccaaag cgcagattct ggaaagagaa 660

ggtcgcctgc cggatgccgt tatcgcctgt gttggcggcg gttcgaatgc catcggcatg 720

tttgctgatt tcatcaatga aaccaacgtc ggcctgattg gtgtggagcc aggtggtcac 780

ggtatcgaaa ctggcgagca cggcgcaccg ctaaaacatg gtcgcgtggg tatctatttc 840

ggtatgaaag cgccgatgat gcaaaccgaa gacgggcaga ttgaagaatc ttactccatc 900

tccgccggac tggatttccc gtctgtcggc ccacaacacg cgtatcttaa cagcactgga 960

cgcgctgatt acgtgtctat taccgatgat gaagcccttg aagccttcaa aacgctgtgc 1020

ctgcacgaag ggatcatccc ggcgctggaa tcctcccacg ccctggccca tgcgttgaaa 1080

atgatgcgcg aaaacccgga taaagagcag ctactggtgg ttaacctttc cggtcgcggc 1140

gataaagaca tcttcaccgt tcacgatatt ttgaaagcac gaggggaaat ctga 1194

<210> 14

<211> 397

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 14

Met Thr Thr Leu Leu Asn Pro Tyr Phe Gly Glu Phe Gly Gly Met Tyr

1 5 10 15

Val Pro Gln Ile Leu Met Pro Ala Leu Arg Gln Leu Glu Glu Ala Phe

20 25 30

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

35 40 45

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

50 55 60

Ile Thr Ala Gly Thr Asn Thr Thr Leu Tyr Leu Lys Arg Glu Asp Leu

65 70 75 80

Leu His Gly Gly Ala His Lys Thr Asn Gln Val Leu Gly Gln Ala Leu

85 90 95

Leu Ala Lys Arg Met Gly Lys Thr Glu Ile Ile Ala Glu Thr Gly Ala

100 105 110

Gly Gln His Gly Val Ala Ser Ala Leu Ala Ser Ala Leu Leu Gly Leu

115 120 125

Lys Cys Arg Ile Tyr Met Gly Ala Lys Asp Val Glu Arg Gln Ser Pro

130 135 140

Asn Val Phe Arg Met Arg Leu Met Gly Ala Glu Val Ile Pro Val His

145 150 155 160

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

165 170 175

Trp Ser Gly Ser Tyr Glu Thr Ala His Tyr Met Leu Gly Thr Ala Ala

180 185 190

Gly Pro His Pro Tyr Pro Thr Ile Val Arg Glu Phe Gln Arg Met Ile

195 200 205

Gly Glu Glu Thr Lys Ala Gln Ile Leu Glu Arg Glu Gly Arg Leu Pro

210 215 220

Asp Ala Val Ile Ala Cys Val Gly Gly Gly Ser Asn Ala Ile Gly Met

225 230 235 240

Phe Ala Asp Phe Ile Asn Glu Thr Asn Val Gly Leu Ile Gly Val Glu

245 250 255

Pro Gly Gly His Gly Ile Glu Thr Gly Glu His Gly Ala Pro Leu Lys

260 265 270

His Gly Arg Val Gly Ile Tyr Phe Gly Met Lys Ala Pro Met Met Gln

275 280 285

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

290 295 300

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

305 310 315 320

Arg Ala Asp Tyr Val Ser Ile Thr Asp Asp Glu Ala Leu Glu Ala Phe

325 330 335

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

340 345 350

His Ala Leu Ala His Ala Leu Lys Met Met Arg Glu Asn Pro Asp Lys

355 360 365

Glu Gln Leu Leu Val Val Asn Leu Ser Gly Arg Gly Asp Lys Asp Ile

370 375 380

Phe Thr Val His Asp Ile Leu Lys Ala Arg Gly Glu Ile

385 390 395

<210> 15

<211> 1362

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 15

atgatgcaaa ccgttttagc gaaaatcgtc gcagacaagg cgatttgggt agaagcccgc 60

aaacagcagc aaccgctggc cagttttcag aatgaggttc agccgagcac gcgacatttt 120

tatgatgcgc tacagggtgc gcgcacggcg tttattctgg agtgcaagaa agcgtcgccg 180

tcaaaaggcg tgatccgtga tgatttcgat ccagcacgca ttgccgccat ttataaacat 240

tacgcttcgg caatttcggt gctgactgat gagaaatatt ttcaggggag ctttaatttc 300

ctccccatcg tcagccaaat cgccccgcag ccgattttat gtaaagactt cattatcgac 360

ccttaccaga tctatctggc gcgctattac caggccgatg cctgcttatt aatgctttca 420

gtactggatg acgaccaata tcgccagctt gccgccgtcg ctcacagtct ggagatgggg 480

gtgctgaccg aagtcagtaa tgaagaggaa caggagcgcg ccattgcatt gggagcaaag 540

gtcgttggca tcaacaaccg cgatctgcgt gatttgtcga ttgatctcaa ccgtacccgc 600

gagcttgcgc cgaaactggg gcacaacgtg acggtaatca gcgaatccgg catcaatact 660

tacgctcagg tgcgcgagtt aagccacttc gctaacggtt ttctgattgg ttcggcgttg 720

atggcccatg acgatttgca cgccgccgtg cgccgggtgt tgctgggtga gaataaagta 780

tgtggcctga cgcgtgggca agatgctaaa gcagcttatg acgcgggcgc gatttacggt 840

gggttgattt ttgttgcgac atcaccgcgt tgcgtcaacg ttgaacaggc gcaggaagtg 900

atggctgcgg caccgttgca gtatgttggc gtgttccgca atcacgatat tgccgatgtg 960

gtggacaaag ctaaggtgtt atcgctggcg gcagtgcaac tgcatggtaa tgaagaacag 1020

ctgtatatcg atacgctgcg tgaagctctg ccagcacatg ttgccatctg gaaagcatta 1080

agcgtcggtg aaaccctgcc cgcccgcgag tttcagcacg ttgataaata tgttttagac 1140

aacggccagg gtggaagcgg gcaacgtttt gactggtcac tattaaatgg tcaatcgctt 1200

ggcaacgttc tgctggcggg gggcttaggc gcagataact gcgtggaagc ggcacaaacc 1260

ggctgcgccg gacttgattt taattctgct gtagagtcgc aaccgggcat caaagacgca 1320

cgtcttttgg cctcggtttt ccagacgctg cgcgcatatt aa 1362

<210> 16

<211> 453

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 16

Met Met Gln Thr Val Leu Ala Lys Ile Val Ala Asp Lys Ala Ile Trp

1 5 10 15

Val Glu Ala Arg Lys Gln Gln Gln Pro Leu Ala Ser Phe Gln Asn Glu

20 25 30

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

35 40 45

Thr Ala Phe Ile Leu Glu Cys Lys Lys Ala Ser Pro Ser Lys Gly Val

50 55 60

Ile Arg Asp Asp Phe Asp Pro Ala Arg Ile Ala Ala Ile Tyr Lys His

65 70 75 80

Tyr Ala Ser Ala Ile Ser Val Leu Thr Asp Glu Lys Tyr Phe Gln Gly

85 90 95

Ser Phe Asn Phe Leu Pro Ile Val Ser Gln Ile Ala Pro Gln Pro Ile

100 105 110

Leu Cys Lys Asp Phe Ile Ile Asp Pro Tyr Gln Ile Tyr Leu Ala Arg

115 120 125

Tyr Tyr Gln Ala Asp Ala Cys Leu Leu Met Leu Ser Val Leu Asp Asp

130 135 140

Asp Gln Tyr Arg Gln Leu Ala Ala Val Ala His Ser Leu Glu Met Gly

145 150 155 160

Val Leu Thr Glu Val Ser Asn Glu Glu Glu Gln Glu Arg Ala Ile Ala

165 170 175

Leu Gly Ala Lys Val Val Gly Ile Asn Asn Arg Asp Leu Arg Asp Leu

180 185 190

Ser Ile Asp Leu Asn Arg Thr Arg Glu Leu Ala Pro Lys Leu Gly His

195 200 205

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

210 215 220

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

225 230 235 240

Met Ala His Asp Asp Leu His Ala Ala Val Arg Arg Val Leu Leu Gly

245 250 255

Glu Asn Lys Val Cys Gly Leu Thr Arg Gly Gln Asp Ala Lys Ala Ala

260 265 270

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

275 280 285

Pro Arg Cys Val Asn Val Glu Gln Ala Gln Glu Val Met Ala Ala Ala

290 295 300

Pro Leu Gln Tyr Val Gly Val Phe Arg Asn His Asp Ile Ala Asp Val

305 310 315 320

Val Asp Lys Ala Lys Val Leu Ser Leu Ala Ala Val Gln Leu His Gly

325 330 335

Asn Glu Glu Gln Leu Tyr Ile Asp Thr Leu Arg Glu Ala Leu Pro Ala

340 345 350

His Val Ala Ile Trp Lys Ala Leu Ser Val Gly Glu Thr Leu Pro Ala

355 360 365

Arg Glu Phe Gln His Val Asp Lys Tyr Val Leu Asp Asn Gly Gln Gly

370 375 380

Gly Ser Gly Gln Arg Phe Asp Trp Ser Leu Leu Asn Gly Gln Ser Leu

385 390 395 400

Gly Asn Val Leu Leu Ala Gly Gly Leu Gly Ala Asp Asn Cys Val Glu

405 410 415

Ala Ala Gln Thr Gly Cys Ala Gly Leu Asp Phe Asn Ser Ala Val Glu

420 425 430

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

435 440 445

Thr Leu Arg Ala Tyr

450

<210> 17

<211> 1596

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 17

atggctgaca ttctgctgct cgataatatc gactctttta cgtacaacct ggcagatcag 60

ttgcgcagca atgggcataa cgtggtgatt taccgcaacc atattccggc gcaaacctta 120

attgaacgcc tggcgaccat gagcaatccg gtgctgatgc tttctcctgg ccccggtgtg 180

ccgagcgaag ccggttgtat gccggaactc ctcacccgct tgcgtggcaa gctgcccatt 240

attggcattt gcctcggaca tcaggcgatt gtcgaagctt acgggggcta tgtcggtcag 300

gcgggcgaaa ttctccacgg taaagcctcc agcattgaac atgacggtca ggcgatgttt 360

gccggattaa caaacccgct gccggtggcg cgttatcact cgctggttgg cagtaacatt 420

ccggccggtt taaccatcaa cgcccatttt aatggcatgg tgatggcagt acgtcacgat 480

gcggatcgcg tttgtggatt ccagttccat ccggaatcca ttctcaccac ccagggcgct 540

cgcctgctgg aacaaacgct ggcctgggcg cagcagaaac tagagccagc caacacgctg 600

caaccgattc tggaaaaact gtatcaggcg cagacgctta gccaacaaga aagccaccag 660

ctgttttcag cggtggtgcg tggcgagctg aagccggaac aactggcggc ggcgctggtg 720

agcatgaaaa ttcgcggtga gcacccgaac gagatcgccg gggcagcaac cgcgctactg 780

gaaaacgcag cgccgttccc gcgcccggat tatctgtttg ctgatatcgt cggtactggc 840

ggtgacggca gcaacagtat caatatttct accgccagtg cgtttgtcgc cgcggcctgt 900

gggctgaaag tggcgaaaca cggcaaccgt agcgtctcca gtaaatctgg ttcgtccgat 960

ctgctggcgg cgttcggtat taatcttgat atgaacgccg ataaatcgcg ccaggcgctg 1020

gatgagttag gtgtatgttt cctctttgcg ccgaagtatc acaccggatt ccgccacgcg 1080

atgccggttc gccagcaact gaaaacccgc accctgttca atgtgctggg gccattgatt 1140

aacccggcgc atccgccgct ggcgttaatt ggtgtttata gtccggaact ggtgctgccg 1200

attgccgaaa ccttgcgcgt gctggggtat caacgcgcgg cggtggtgca cagcggcggg 1260

atggatgaag tttcattaca cgcgccgaca atcgttgccg aactgcatga cggcgaaatt 1320

aaaagctatc agctcaccgc agaagacttt ggcctgacac cctaccacca ggagcaactg 1380

gcaggcggaa caccggaaga aaaccgtgac attttaacac gtttgttaca aggtaaaggc 1440

gacgccgccc atgaagcagc cgtcgctgcg aacgtcgcca tgttaatgcg cctgcatggc 1500

catgaagatc tgcaagccaa tgcgcaaacc gttcttgagg tactgcgcag tggttccgct 1560

tacgacagag tcaccgcact ggcggcacga gggtaa 1596

<210> 18

<211> 531

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 18

Met Ala Asp Ile Leu Leu Leu Asp Asn Ile Asp Ser Phe Thr Tyr Asn

1 5 10 15

Leu Ala Asp Gln Leu Arg Ser Asn Gly His Asn Val Val Ile Tyr Arg

20 25 30

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

35 40 45

Asn Pro Val Leu Met Leu Ser Pro Gly Pro Gly Val Pro Ser Glu Ala

50 55 60

Gly Cys Met Pro Glu Leu Leu Thr Arg Leu Arg Gly Lys Leu Pro Ile

65 70 75 80

Ile Gly Ile Cys Leu Gly His Gln Ala Ile Val Glu Ala Tyr Gly Gly

85 90 95

Tyr Val Gly Gln Ala Gly Glu Ile Leu His Gly Lys Ala Ser Ser Ile

100 105 110

Glu His Asp Gly Gln Ala Met Phe Ala Gly Leu Thr Asn Pro Leu Pro

115 120 125

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

130 135 140

Thr Ile Asn Ala His Phe Asn Gly Met Val Met Ala Val Arg His Asp

145 150 155 160

Ala Asp Arg Val Cys Gly Phe Gln Phe His Pro Glu Ser Ile Leu Thr

165 170 175

Thr Gln Gly Ala Arg Leu Leu Glu Gln Thr Leu Ala Trp Ala Gln Gln

180 185 190

Lys Leu Glu Pro Ala Asn Thr Leu Gln Pro Ile Leu Glu Lys Leu Tyr

195 200 205

Gln Ala Gln Thr Leu Ser Gln Gln Glu Ser His Gln Leu Phe Ser Ala

210 215 220

Val Val Arg Gly Glu Leu Lys Pro Glu Gln Leu Ala Ala Ala Leu Val

225 230 235 240

Ser Met Lys Ile Arg Gly Glu His Pro Asn Glu Ile Ala Gly Ala Ala

245 250 255

Thr Ala Leu Leu Glu Asn Ala Ala Pro Phe Pro Arg Pro Asp Tyr Leu

260 265 270

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

275 280 285

Ile Ser Thr Ala Ser Ala Phe Val Ala Ala Ala Cys Gly Leu Lys Val

290 295 300

Ala Lys His Gly Asn Arg Ser Val Ser Ser Lys Ser Gly Ser Ser Asp

305 310 315 320

Leu Leu Ala Ala Phe Gly Ile Asn Leu Asp Met Asn Ala Asp Lys Ser

325 330 335

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

340 345 350

Tyr His Thr Gly Phe Arg His Ala Met Pro Val Arg Gln Gln Leu Lys

355 360 365

Thr Arg Thr Leu Phe Asn Val Leu Gly Pro Leu Ile Asn Pro Ala His

370 375 380

Pro Pro Leu Ala Leu Ile Gly Val Tyr Ser Pro Glu Leu Val Leu Pro

385 390 395 400

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

405 410 415

His Ser Gly Gly Met Asp Glu Val Ser Leu His Ala Pro Thr Ile Val

420 425 430

Ala Glu Leu His Asp Gly Glu Ile Lys Ser Tyr Gln Leu Thr Ala Glu

435 440 445

Asp Phe Gly Leu Thr Pro Tyr His Gln Glu Gln Leu Ala Gly Gly Thr

450 455 460

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

465 470 475 480

Asp Ala Ala His Glu Ala Ala Val Ala Ala Asn Val Ala Met Leu Met

485 490 495

Arg Leu His Gly His Glu Asp Leu Gln Ala Asn Ala Gln Thr Val Leu

500 505 510

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

515 520 525

Ala Arg Gly

530

<210> 19

<211> 1908

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 19

atggcacctg ttacaattga aaagttcgta aatcaagaag aacgacacct tgtttccaac 60

cgatcagcaa caattccgtt tggtgaatac atatttaaaa gattgttgtc catcgatacg 120

aaatcagttt tcggtgttcc tggtgacttc aacttatctc tattagaata tctctattca 180

cctagtgttg aatcagctgg cctaagatgg gtcggcacgt gtaatgaact gaacgccgct 240

tatgcggccg acggatattc ccgttactct aataagattg gctgtttaat aaccacgtat 300

ggcgttggtg aattaagcgc cttgaacggt atagccggtt cgttcgctga aaatgtcaaa 360

gttttgcaca ttgttggtgt ggccaagtcc atagattcgc gttcaagtaa ctttagtgat 420

cggaacctac atcatttggt cccacagcta catgattcaa attttaaagg gccaaatcat 480

aaagtatatc atgatatggt aaaagataga gtcgcttgct cggtagccta cttggaggat 540

attgaaactg catgtgacca agtcgataat gttatccgcg atatttacaa gtattctaaa 600

cctggttata tttttgttcc tgcagatttt gcggatatgt ctgttacatg tgataatttg 660

gttaatgttc cacgtatatc tcaacaagat tgtatagtat acccttctga aaaccaattg 720

tctgacataa tcaacaagat tactagttgg atatattcca gtaaaacacc tgcgatcctt 780

ggagacgtac tgactgatag gtatggtgtg agtaactttt tgaacaagct tatctgcaaa 840

actgggattt ggaatttttc cactgttatg ggaaaatctg taattgatga gtcaaaccca 900

acttatatgg gtcaatataa tggtaaagaa ggtttaaaac aagtctatga acattttgaa 960

ctgtgcgact tggtcttgca ttttggagtc gacatcaatg aaattaataa tgggcattat 1020

acttttactt ataaaccaaa tgctaaaatc attcaatttc atccgaatta tattcgcctt 1080

gtggacacta ggcagggcaa tgagcaaatg ttcaaaggaa tcaattttgc ccctatttta 1140

aaagaactat acaagcgcat tgacgtttct aaactttctt tgcaatatga ttcaaatgta 1200

actcaatata cgaacgaaac aatgcggtta gaagatccta ccaatggaca atcaagcatt 1260

attacacaag ttcacttaca aaagacgatg cctaaatttt tgaaccctgg tgatgttgtc 1320

gtttgtgaaa caggctcttt tcaattctct gttcgtgatt tcgcgtttcc ttcgcaatta 1380

aaatatatat cgcaaggatt tttcctttcc attggcatgg cccttcctgc cgccctaggt 1440

gttggaattg ccatgcaaga ccactcaaac gctcacatca atggtggcaa cgtaaaagag 1500

gactataagc caagattaat tttgtttgaa ggtgacggtg cagcacagat gacaatccaa 1560

gaactgagca ccattctgaa gtgcaatatt ccactagaag ttatcatttg gaacaataac 1620

ggctacacta ttgaaagagc catcatgggc cctaccaggt cgtataacga cgttatgtct 1680

tggaaatgga ccaaactatt tgaagcattc ggagacttcg acggaaagta tactaatagc 1740

actctcattc aatgtccctc taaattagca ctgaaattgg aggagcttaa gaattcaaac 1800

aaaagaagcg ggatagaact tttagaagtc aaattaggcg aattggattt ccccgaacag 1860

ctaaagtgca tggttgaagc agcggcactt aaaagaaata aaaaatag 1908

<210> 20

<211> 635

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 20

Met Ala Pro Val Thr Ile Glu Lys Phe Val Asn Gln Glu Glu Arg His

1 5 10 15

Leu Val Ser Asn Arg Ser Ala Thr Ile Pro Phe Gly Glu Tyr Ile Phe

20 25 30

Lys Arg Leu Leu Ser Ile Asp Thr Lys Ser Val Phe Gly Val Pro Gly

35 40 45

Asp Phe Asn Leu Ser Leu Leu Glu Tyr Leu Tyr Ser Pro Ser Val Glu

50 55 60

Ser Ala Gly Leu Arg Trp Val Gly Thr Cys Asn Glu Leu Asn Ala Ala

65 70 75 80

Tyr Ala Ala Asp Gly Tyr Ser Arg Tyr Ser Asn Lys Ile Gly Cys Leu

85 90 95

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

100 105 110

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

115 120 125

Lys Ser Ile Asp Ser Arg Ser Ser Asn Phe Ser Asp Arg Asn Leu His

130 135 140

His Leu Val Pro Gln Leu His Asp Ser Asn Phe Lys Gly Pro Asn His

145 150 155 160

Lys Val Tyr His Asp Met Val Lys Asp Arg Val Ala Cys Ser Val Ala

165 170 175

Tyr Leu Glu Asp Ile Glu Thr Ala Cys Asp Gln Val Asp Asn Val Ile

180 185 190

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

195 200 205

Asp Phe Ala Asp Met Ser Val Thr Cys Asp Asn Leu Val Asn Val Pro

210 215 220

Arg Ile Ser Gln Gln Asp Cys Ile Val Tyr Pro Ser Glu Asn Gln Leu

225 230 235 240

Ser Asp Ile Ile Asn Lys Ile Thr Ser Trp Ile Tyr Ser Ser Lys Thr

245 250 255

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

260 265 270

Phe Leu Asn Lys Leu Ile Cys Lys Thr Gly Ile Trp Asn Phe Ser Thr

275 280 285

Val Met Gly Lys Ser Val Ile Asp Glu Ser Asn Pro Thr Tyr Met Gly

290 295 300

Gln Tyr Asn Gly Lys Glu Gly Leu Lys Gln Val Tyr Glu His Phe Glu

305 310 315 320

Leu Cys Asp Leu Val Leu His Phe Gly Val Asp Ile Asn Glu Ile Asn

325 330 335

Asn Gly His Tyr Thr Phe Thr Tyr Lys Pro Asn Ala Lys Ile Ile Gln

340 345 350

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

355 360 365

Gln Met Phe Lys Gly Ile Asn Phe Ala Pro Ile Leu Lys Glu Leu Tyr

370 375 380

Lys Arg Ile Asp Val Ser Lys Leu Ser Leu Gln Tyr Asp Ser Asn Val

385 390 395 400

Thr Gln Tyr Thr Asn Glu Thr Met Arg Leu Glu Asp Pro Thr Asn Gly

405 410 415

Gln Ser Ser Ile Ile Thr Gln Val His Leu Gln Lys Thr Met Pro Lys

420 425 430

Phe Leu Asn Pro Gly Asp Val Val Val Cys Glu Thr Gly Ser Phe Gln

435 440 445

Phe Ser Val Arg Asp Phe Ala Phe Pro Ser Gln Leu Lys Tyr Ile Ser

450 455 460

Gln Gly Phe Phe Leu Ser Ile Gly Met Ala Leu Pro Ala Ala Leu Gly

465 470 475 480

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

485 490 495

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

500 505 510

Gly Ala Ala Gln Met Thr Ile Gln Glu Leu Ser Thr Ile Leu Lys Cys

515 520 525

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

530 535 540

Glu Arg Ala Ile Met Gly Pro Thr Arg Ser Tyr Asn Asp Val Met Ser

545 550 555 560

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

565 570 575

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

580 585 590

Leu Glu Glu Leu Lys Asn Ser Asn Lys Arg Ser Gly Ile Glu Leu Leu

595 600 605

Glu Val Lys Leu Gly Glu Leu Asp Phe Pro Glu Gln Leu Lys Cys Met

610 615 620

Val Glu Ala Ala Ala Leu Lys Arg Asn Lys Lys

625 630 635

<210> 21

<211> 1488

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 21

atgaattttc atcatctggc ttactggcag gataaagcgt taagtctcgc cattgaaaac 60

cgcttattta ttaacggtga atatactgct gcggcggaaa atgaaacctt tgaaaccgtt 120

gatccggtca cccaggcacc gctggcgaaa attgcccgcg gcaagagcgt cgatatcgac 180

cgtgcgatga gcgcagcacg cggcgtattt gaacgcggcg actggtcact ctcttctccg 240

gctaaacgta aagcggtact gaataaactc gccgatttaa tggaagccca cgccgaagag 300

ctggcactgc tggaaactct cgacaccggc aaaccgattc gtcacagtct gcgtgatgat 360

attcccggcg cggcgcgcgc cattcgctgg tacgccgaag cgatcgacaa agtgtatggc 420

gaagtggcga ccaccagtag ccatgagctg gcgatgatcg tgcgtgaacc ggtcggcgtg 480

attgccgcca tcgtgccgtg gaacttcccg ctgttgctga cttgctggaa actcggcccg 540

gcgctggcgg cgggaaacag cgtgattcta aaaccgtctg aaaaatcacc gctcagtgcg 600

attcgtctcg cggggctggc gaaagaagca ggcttgccgg atggtgtgtt gaacgtggtg 660

acgggttttg gtcatgaagc cgggcaggcg ctgtcgcgtc ataacgatat cgacgccatt 720

gcctttaccg gttcaacccg taccgggaaa cagctgctga aagatgcggg cgacagcaac 780

atgaaacgcg tctggctgga agcgggcggc aaaagcgcca acatcgtttt cgctgactgc 840

ccggatttgc aacaggcggc aagcgccacc gcagcaggca ttttctacaa ccagggacag 900

gtgtgcatcg ccggaacgcg cctgttgctg gaagagagca tcgccgatga attcttagcc 960

ctgttaaaac agcaggcgca aaactggcag ccgggccatc cacttgatcc cgcaaccacc 1020

atgggcacct taatcgactg cgcccacgcc gactcggtcc atagctttat tcgggaaggc 1080

gaaagcaaag ggcaactgtt gttggatggc cgtaacgccg ggctggctgc cgccatcggc 1140

ccgaccatct ttgtggatgt ggacccgaat gcgtccttaa gtcgcgaaga gattttcggt 1200

ccggtgctgg tggtcacgcg tttcacatca gaagaacagg cgctacagct tgccaacgac 1260

agccagtacg gccttggcgc ggcggtatgg acgcgcgacc tctcccgcgc gcaccgcatg 1320

agccgacgcc tgaaagccgg ttccgtcttc gtcaataact acaacgacgg cgatatgacc 1380

gtgccgtttg gcggctataa gcagagcggc aacggtcgcg acaaatccct gcatgccctt 1440

gaaaaattca ctgaactgaa aaccatctgg ataagcctgg aggcctga 1488

<210> 22

<211> 495

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 22

Met Asn Phe His His Leu Ala Tyr Trp Gln Asp Lys Ala Leu Ser Leu

1 5 10 15

Ala Ile Glu Asn Arg Leu Phe Ile Asn Gly Glu Tyr Thr Ala Ala Ala

20 25 30

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

35 40 45

Ala Lys Ile Ala Arg Gly Lys Ser Val Asp Ile Asp Arg Ala Met Ser

50 55 60

Ala Ala Arg Gly Val Phe Glu Arg Gly Asp Trp Ser Leu Ser Ser Pro

65 70 75 80

Ala Lys Arg Lys Ala Val Leu Asn Lys Leu Ala Asp Leu Met Glu Ala

85 90 95

His Ala Glu Glu Leu Ala Leu Leu Glu Thr Leu Asp Thr Gly Lys Pro

100 105 110

Ile Arg His Ser Leu Arg Asp Asp Ile Pro Gly Ala Ala Arg Ala Ile

115 120 125

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

130 135 140

Thr Ser Ser His Glu Leu Ala Met Ile Val Arg Glu Pro Val Gly Val

145 150 155 160

Ile Ala Ala Ile Val Pro Trp Asn Phe Pro Leu Leu Leu Thr Cys Trp

165 170 175

Lys Leu Gly Pro Ala Leu Ala Ala Gly Asn Ser Val Ile Leu Lys Pro

180 185 190

Ser Glu Lys Ser Pro Leu Ser Ala Ile Arg Leu Ala Gly Leu Ala Lys

195 200 205

Glu Ala Gly Leu Pro Asp Gly Val Leu Asn Val Val Thr Gly Phe Gly

210 215 220

His Glu Ala Gly Gln Ala Leu Ser Arg His Asn Asp Ile Asp Ala Ile

225 230 235 240

Ala Phe Thr Gly Ser Thr Arg Thr Gly Lys Gln Leu Leu Lys Asp Ala

245 250 255

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

260 265 270

Ala Asn Ile Val Phe Ala Asp Cys Pro Asp Leu Gln Gln Ala Ala Ser

275 280 285

Ala Thr Ala Ala Gly Ile Phe Tyr Asn Gln Gly Gln Val Cys Ile Ala

290 295 300

Gly Thr Arg Leu Leu Leu Glu Glu Ser Ile Ala Asp Glu Phe Leu Ala

305 310 315 320

Leu Leu Lys Gln Gln Ala Gln Asn Trp Gln Pro Gly His Pro Leu Asp

325 330 335

Pro Ala Thr Thr Met Gly Thr Leu Ile Asp Cys Ala His Ala Asp Ser

340 345 350

Val His Ser Phe Ile Arg Glu Gly Glu Ser Lys Gly Gln Leu Leu Leu

355 360 365

Asp Gly Arg Asn Ala Gly Leu Ala Ala Ala Ile Gly Pro Thr Ile Phe

370 375 380

Val Asp Val Asp Pro Asn Ala Ser Leu Ser Arg Glu Glu Ile Phe Gly

385 390 395 400

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

405 410 415

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

420 425 430

Asp Leu Ser Arg Ala His Arg Met Ser Arg Arg Leu Lys Ala Gly Ser

435 440 445

Val Phe Val Asn Asn Tyr Asn Asp Gly Asp Met Thr Val Pro Phe Gly

450 455 460

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

465 470 475 480

Glu Lys Phe Thr Glu Leu Lys Thr Ile Trp Ile Ser Leu Glu Ala

485 490 495

<210> 23

<211> 1503

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 23

atgactttac ctgaatcaaa agacttttct tacttgtttt cggatgaaac caatgctcgt 60

aaaccatccc cattgaaaac ctgcatccat cttttccaag atcctaacat tatctttttg 120

ggtggtggcc tgccattaaa agattatttc ccatgggata atctatctgt agattcaccc 180

aagcctcctt ttccccaggg tattggagct ccaattgacg agcagaattg cataaaatac 240

accgtcaaca aagattacgc tgataaaagt gccaatcctt ccaacgatat tcctttgtca 300

agagctttgc aatacgggtt cagtgctggt caacctgaac tattaaactt cattagagat 360

cataccaaga ttatccacga tttgaagtat aaggactggg acgttttagc cactgcaggt 420

aacacaaatg cctgggaatc tactttaaga gtcttttgta accgaggtga tgtcatctta 480

gttgaggcac attctttttc ctcttcattg gcttctgcag aggctcaagg tgtcattacc 540

ttccccgtgc caattgacgc tgatggtatc attcctgaaa aattagctaa agtcatggaa 600

aactggacac ctggtgctcc taaaccaaag ttgttataca ctattccaac gggccaaaat 660

ccaactggta cttccattgc agaccataga aaggaggcaa tttacaagat cgctcaaaag 720

tacgacttcc taattgtgga agatgaacct tattatttct tacaaatgaa tccctacatc 780

aaagacttga aggaaagaga gaaggcacaa agttctccaa agcaggacca tgacgaattt 840

ttgaagtcct tggcaaacac tttcctttcc ttggatacag aaggccgtgt tattagaatg 900

gattcctttt caaaagtttt ggccccaggg acaagattgg gttggattac tggttcatcc 960

aaaatcttga agccttactt gagtttgcat gaaatgacga ttcaagcccc agcaggtttt 1020

acacaagttt tggtcaacgc tacgctatcc aggtggggtc aaaagggtta cttggactgg 1080

ttgcttggcc tgcgtcatga atacactttg aaacgtgact gtgccatcga tgccctttac 1140

aagtatctac cacaatctga tgctttcgtg atcaatcctc caattgcagg tatgtttttc 1200

accgtgaaca ttgacgcatc tgtccaccct gagtttaaaa caaaatacaa ctcagaccct 1260

taccagctag aacagagtct ttaccacaaa gtggttgaac gtggtgtttt agtggttccc 1320

ggttcttggt tcaagagtga gggtgagacg gaacctcctc aacccgctga atctaaagaa 1380

gtcagtaatc caaacataat tttcttcaga ggtacctatg cagctgtctc tcctgagaaa 1440

ctgactgaag gtctgaagag attaggtgat actttatacg aagaatttgg tatttccaaa 1500

tag 1503

<210> 24

<211> 500

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 24

Met Thr Leu Pro Glu Ser Lys Asp Phe Ser Tyr Leu Phe Ser Asp Glu

1 5 10 15

Thr Asn Ala Arg Lys Pro Ser Pro Leu Lys Thr Cys Ile His Leu Phe

20 25 30

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

35 40 45

Tyr Phe Pro Trp Asp Asn Leu Ser Val Asp Ser Pro Lys Pro Pro Phe

50 55 60

Pro Gln Gly Ile Gly Ala Pro Ile Asp Glu Gln Asn Cys Ile Lys Tyr

65 70 75 80

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

85 90 95

Ile Pro Leu Ser Arg Ala Leu Gln Tyr Gly Phe Ser Ala Gly Gln Pro

100 105 110

Glu Leu Leu Asn Phe Ile Arg Asp His Thr Lys Ile Ile His Asp Leu

115 120 125

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

130 135 140

Trp Glu Ser Thr Leu Arg Val Phe Cys Asn Arg Gly Asp Val Ile Leu

145 150 155 160

Val Glu Ala His Ser Phe Ser Ser Ser Leu Ala Ser Ala Glu Ala Gln

165 170 175

Gly Val Ile Thr Phe Pro Val Pro Ile Asp Ala Asp Gly Ile Ile Pro

180 185 190

Glu Lys Leu Ala Lys Val Met Glu Asn Trp Thr Pro Gly Ala Pro Lys

195 200 205

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

210 215 220

Ser Ile Ala Asp His Arg Lys Glu Ala Ile Tyr Lys Ile Ala Gln Lys

225 230 235 240

Tyr Asp Phe Leu Ile Val Glu Asp Glu Pro Tyr Tyr Phe Leu Gln Met

245 250 255

Asn Pro Tyr Ile Lys Asp Leu Lys Glu Arg Glu Lys Ala Gln Ser Ser

260 265 270

Pro Lys Gln Asp His Asp Glu Phe Leu Lys Ser Leu Ala Asn Thr Phe

275 280 285

Leu Ser Leu Asp Thr Glu Gly Arg Val Ile Arg Met Asp Ser Phe Ser

290 295 300

Lys Val Leu Ala Pro Gly Thr Arg Leu Gly Trp Ile Thr Gly Ser Ser

305 310 315 320

Lys Ile Leu Lys Pro Tyr Leu Ser Leu His Glu Met Thr Ile Gln Ala

325 330 335

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

340 345 350

Gly Gln Lys Gly Tyr Leu Asp Trp Leu Leu Gly Leu Arg His Glu Tyr

355 360 365

Thr Leu Lys Arg Asp Cys Ala Ile Asp Ala Leu Tyr Lys Tyr Leu Pro

370 375 380

Gln Ser Asp Ala Phe Val Ile Asn Pro Pro Ile Ala Gly Met Phe Phe

385 390 395 400

Thr Val Asn Ile Asp Ala Ser Val His Pro Glu Phe Lys Thr Lys Tyr

405 410 415

Asn Ser Asp Pro Tyr Gln Leu Glu Gln Ser Leu Tyr His Lys Val Val

420 425 430

Glu Arg Gly Val Leu Val Val Pro Gly Ser Trp Phe Lys Ser Glu Gly

435 440 445

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

450 455 460

Asn Ile Ile Phe Phe Arg Gly Thr Tyr Ala Ala Val Ser Pro Glu Lys

465 470 475 480

Leu Thr Glu Gly Leu Lys Arg Leu Gly Asp Thr Leu Tyr Glu Glu Phe

485 490 495

Gly Ile Ser Lys

500

<210> 25

<211> 327

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 25

atggcccaac aatcacccta ttcagcagcg atggcagaac agcgtcacca ggagtggtta 60

cgttttgtcg acctgcttaa gaatgcctac caaaacgatc tccatttacc gttgttaaac 120

ctgatgctga cgccagatga gcgcgaagcg ttggggactc gcgtgcgtat tgtcgaagag 180

ctgttgcgcg gcgaaatgag ccagcgtgag ttaaaaaatg aactcggcgc aggcatcgcg 240

acgattacgc gtggatctaa cagcctgaaa gccgcgcccg tcgagctgcg ccagtggctg 300

gaagaggtgt tgctgaaaag cgattga 327

<210> 26

<211> 108

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 26

Met Ala Gln Gln Ser Pro Tyr Ser Ala Ala Met Ala Glu Gln Arg His

1 5 10 15

Gln Glu Trp Leu Arg Phe Val Asp Leu Leu Lys Asn Ala Tyr Gln Asn

20 25 30

Asp Leu His Leu Pro Leu Leu Asn Leu Met Leu Thr Pro Asp Glu Arg

35 40 45

Glu Ala Leu Gly Thr Arg Val Arg Ile Val Glu Glu Leu Leu Arg Gly

50 55 60

Glu Met Ser Gln Arg Glu Leu Lys Asn Glu Leu Gly Ala Gly Ile Ala

65 70 75 80

Thr Ile Thr Arg Gly Ser Asn Ser Leu Lys Ala Ala Pro Val Glu Leu

85 90 95

Arg Gln Trp Leu Glu Glu Val Leu Leu Lys Ser Asp

100 105

<210> 27

<211> 804

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 27

atggctatcc ctgcatttgg tttaggtact ttccgtctga aagacgacgt tgttatttca 60

tctgtgataa cggcgcttga acttggttat cgcgcaattg ataccgcaca aatctatgat 120

aacgaagccg cagtaggtca ggcgattgca gaaagtggcg tgccacgtca tgaactctac 180

atcaccacta aaatctggat tgaaaatctc agcaaagaca aattgatccc aagtctgaaa 240

gagagcctgc aaaaattgcg taccgattat gttgatctga cgctaatcca ctggccgtca 300

ccaaacgatg aagtctctgt tgaagagttt atgcaggcgc tgctggaagc caaaaaacaa 360

gggctgacgc gtgagatcgg tatttccaac ttcacgatcc cgttgatgga aaaagcgatt 420

gctgctgttg gtgctgaaaa catcgctact aaccagattg aactctctcc ttatctgcaa 480

aaccgtaaag tggttgcctg ggctaaacag cacggcatcc atattacttc ctatatgacg 540

ctggcgtatg gtaaggccct gaaagatgag gttattgctc gtatcgcagc taaacacaat 600

gcgactccgg cacaagtgat tctggcgtgg gctatggggg aaggttactc agtaattcct 660

tcttctacta aacgtaaaaa cctggaaagt aatcttaagg cacaaaattt acagcttgat 720

gccgaagata aaaaagcgat cgccgcactg gattgcaacg accgcctggt tagcccggaa 780

ggtctggctc ctgaatggga ttaa 804

<210> 28

<211> 267

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 28

Met Ala Ile Pro Ala Phe Gly Leu Gly Thr Phe Arg Leu Lys Asp Asp

1 5 10 15

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

20 25 30

Ile Asp Thr Ala Gln Ile Tyr Asp Asn Glu Ala Ala Val Gly Gln Ala

35 40 45

Ile Ala Glu Ser Gly Val Pro Arg His Glu Leu Tyr Ile Thr Thr Lys

50 55 60

Ile Trp Ile Glu Asn Leu Ser Lys Asp Lys Leu Ile Pro Ser Leu Lys

65 70 75 80

Glu Ser Leu Gln Lys Leu Arg Thr Asp Tyr Val Asp Leu Thr Leu Ile

85 90 95

His Trp Pro Ser Pro Asn Asp Glu Val Ser Val Glu Glu Phe Met Gln

100 105 110

Ala Leu Leu Glu Ala Lys Lys Gln Gly Leu Thr Arg Glu Ile Gly Ile

115 120 125

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

130 135 140

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

145 150 155 160

Asn Arg Lys Val Val Ala Trp Ala Lys Gln His Gly Ile His Ile Thr

165 170 175

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

180 185 190

Ala Arg Ile Ala Ala Lys His Asn Ala Thr Pro Ala Gln Val Ile Leu

195 200 205

Ala Trp Ala Met Gly Glu Gly Tyr Ser Val Ile Pro Ser Ser Thr Lys

210 215 220

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

225 230 235 240

Ala Glu Asp Lys Lys Ala Ile Ala Ala Leu Asp Cys Asn Asp Arg Leu

245 250 255

Val Ser Pro Glu Gly Leu Ala Pro Glu Trp Asp

260 265

<210> 29

<211> 855

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 29

atgcaacaaa aaatgattca atttagtggc gatgtctcac tgccagccgt agggcaggga 60

acatggtata tgggcgaaga tgccagtcag cgcaaaacag aagttgctgc actacgcgcg 120

ggcattgaac tcggtttaac cctcattgat accgccgaaa tgtatgccga tggcggtgcc 180

gaaaaggtgg ttggggaagc attaaccggt ctgcgagaga aggtctttct cgtctctaaa 240

gtctatccgt ggaatgctgg cgggcaaaaa gcgataaatg catgcgaagc cagtttacgc 300

cgtctcaata ctgattatct cgatctttac ttattacact ggtctggcag tttcgctttt 360

gaagagactg tcgcagcgat ggaaaaattg atcgcccagg gaaaaatccg ccgctggggc 420

gtttctaacc ttgattatgc tgatatgcag gaactctggc agctgccggg gggaaatcag 480

tgtgccacta atcaggtgct ttaccatctc ggttcacgag gaattgagta cgatctactc 540

ccctggtgcc agcaacagca gatgccggtg atggcttaca gtccgttagc ccaggccggg 600

cggttgcgca atggactgtt aaaaaacgcg gtagtcaacg aaattgcaca tgctcacaat 660

atcagcgcgg cacaagtatt gttggcgtgg gtgatcagtc atcagggtgt gatggcgatt 720

ccaaaagcgg ccacgattgc ccatgtccaa caaaatgcgg ctgtgcttga ggtcgaactt 780

tcttcagcgg aattagctat gctggataag gcatatccgg caccaaaagg aaaaactgcg 840

ctggatatgg tgtga 855

<210> 30

<211> 284

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 30

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

1 5 10 15

Val Gly Gln Gly Thr Trp Tyr Met Gly Glu Asp Ala Ser Gln Arg Lys

20 25 30

Thr Glu Val Ala Ala Leu Arg Ala Gly Ile Glu Leu Gly Leu Thr Leu

35 40 45

Ile Asp Thr Ala Glu Met Tyr Ala Asp Gly Gly Ala Glu Lys Val Val

50 55 60

Gly Glu Ala Leu Thr Gly Leu Arg Glu Lys Val Phe Leu Val Ser Lys

65 70 75 80

Val Tyr Pro Trp Asn Ala Gly Gly Gln Lys Ala Ile Asn Ala Cys Glu

85 90 95

Ala Ser Leu Arg Arg Leu Asn Thr Asp Tyr Leu Asp Leu Tyr Leu Leu

100 105 110

His Trp Ser Gly Ser Phe Ala Phe Glu Glu Thr Val Ala Ala Met Glu

115 120 125

Lys Leu Ile Ala Gln Gly Lys Ile Arg Arg Trp Gly Val Ser Asn Leu

130 135 140

Asp Tyr Ala Asp Met Gln Glu Leu Trp Gln Leu Pro Gly Gly Asn Gln

145 150 155 160

Cys Ala Thr Asn Gln Val Leu Tyr His Leu Gly Ser Arg Gly Ile Glu

165 170 175

Tyr Asp Leu Leu Pro Trp Cys Gln Gln Gln Gln Met Pro Val Met Ala

180 185 190

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

195 200 205

Asn Ala Val Val Asn Glu Ile Ala His Ala His Asn Ile Ser Ala Ala

210 215 220

Gln Val Leu Leu Ala Trp Val Ile Ser His Gln Gly Val Met Ala Ile

225 230 235 240

Pro Lys Ala Ala Thr Ile Ala His Val Gln Gln Asn Ala Ala Val Leu

245 250 255

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

260 265 270

Pro Ala Pro Lys Gly Lys Thr Ala Leu Asp Met Val

275 280

<210> 31

<211> 957

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 31

atgctacaaa attgcgcaca atcaaattgc cgcattattc ctaagaaatt acgcgatatg 60

aaacgtgaag agatttgccg cttgctggcg gataaagtta ataaactgaa aaataaagaa 120

aatagtttgt caggactgtt gcccgatgtg cgtttgttgt atggcgagac gcctttcgca 180

cgtacaccgg tgatgtacga gcctggcatc ataattctct tttccgggca taaaatcggt 240

tatatcaatg aacgcgtgtt tcgttatgat gccaatgaat acctgctgct gacggtgccg 300

ttgccgtttg agtgcgaaac ctatgccacg tcagaggtgc cgctggcagg gttgcgtctc 360

aatgtcgata ttttgcagtt acaggaactg ttgatggaca ttggcgaaga tgagcatttc 420

cagccgtcga tggcagccag cgggattaac tccgccacgt tatcagaaga gattttatgc 480

gcggcggagc ggttactcga cgtgatggag cgaccactgg atgcgcgtat tctcggcaaa 540

cagatcatcc gcgaaattct gtactacgtg ctgaccggac cttgcggcgg cgcgttactg 600

gcgctggtca gtcgccagac tcacttcagt ctgattagcc gcgtgctgaa acggattgag 660

aataaataca ccgaaaacct gagcgtcgag caactggcgg cagaagccaa catgagcgta 720

tcggcgttcc accataattt taagtctgtc accagtacct cgccgttgca gtatttgaag 780

aattaccgtc tgcataaggc gcggatgatg atcatccatg acggcatgaa ggccagcgca 840

gcagcgatgc gcgtcggcta tgaaagcgca tcgcaattta gccgtgagtt taaacgttac 900

ttcggtgtga cgccggggga agatgcggca agaatgcggg cgatgcaggg gaattaa 957

<210> 32

<211> 318

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 32

Met Leu Gln Asn Cys Ala Gln Ser Asn Cys Arg Ile Ile Pro Lys Lys

1 5 10 15

Leu Arg Asp Met Lys Arg Glu Glu Ile Cys Arg Leu Leu Ala Asp Lys

20 25 30

Val Asn Lys Leu Lys Asn Lys Glu Asn Ser Leu Ser Gly Leu Leu Pro

35 40 45

Asp Val Arg Leu Leu Tyr Gly Glu Thr Pro Phe Ala Arg Thr Pro Val

50 55 60

Met Tyr Glu Pro Gly Ile Ile Ile Leu Phe Ser Gly His Lys Ile Gly

65 70 75 80

Tyr Ile Asn Glu Arg Val Phe Arg Tyr Asp Ala Asn Glu Tyr Leu Leu

85 90 95

Leu Thr Val Pro Leu Pro Phe Glu Cys Glu Thr Tyr Ala Thr Ser Glu

100 105 110

Val Pro Leu Ala Gly Leu Arg Leu Asn Val Asp Ile Leu Gln Leu Gln

115 120 125

Glu Leu Leu Met Asp Ile Gly Glu Asp Glu His Phe Gln Pro Ser Met

130 135 140

Ala Ala Ser Gly Ile Asn Ser Ala Thr Leu Ser Glu Glu Ile Leu Cys

145 150 155 160

Ala Ala Glu Arg Leu Leu Asp Val Met Glu Arg Pro Leu Asp Ala Arg

165 170 175

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

180 185 190

Gly Pro Cys Gly Gly Ala Leu Leu Ala Leu Val Ser Arg Gln Thr His

195 200 205

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

210 215 220

Glu Asn Leu Ser Val Glu Gln Leu Ala Ala Glu Ala Asn Met Ser Val

225 230 235 240

Ser Ala Phe His His Asn Phe Lys Ser Val Thr Ser Thr Ser Pro Leu

245 250 255

Gln Tyr Leu Lys Asn Tyr Arg Leu His Lys Ala Arg Met Met Ile Ile

260 265 270

His Asp Gly Met Lys Ala Ser Ala Ala Ala Met Arg Val Gly Tyr Glu

275 280 285

Ser Ala Ser Gln Phe Ser Arg Glu Phe Lys Arg Tyr Phe Gly Val Thr

290 295 300

Pro Gly Glu Asp Ala Ala Arg Met Arg Ala Met Gln Gly Asn

305 310 315

<210> 33

<211> 1164

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 33

atgaacaact ttaatctgca caccccaacc cgcattctgt ttggtaaagg cgcaatcgct 60

ggtttacgcg aacaaattcc tcacgatgct cgcgtattga ttacctacgg cggcggcagc 120

gtgaaaaaaa ccggcgttct cgatcaagtt ctggatgccc tgaaaggcat ggacgtgctg 180

gaatttggcg gtattgagcc aaacccggct tatgaaacgc tgatgaacgc cgtgaaactg 240

gttcgcgaac agaaagtgac tttcctgctg gcggttggcg gcggttctgt actggacggc 300

accaaattta tcgccgcagc ggctaactat ccggaaaata tcgatccgtg gcacattctg 360

caaacgggcg gtaaagagat taaaagcgcc atcccgatgg gctgtgtgct gacgctgcca 420

gcaaccggtt cagaatccaa cgcaggcgcg gtgatctccc gtaaaaccac aggcgacaag 480

caggcgttcc attctgccca tgttcagccg gtatttgccg tgctcgatcc ggtttatacc 540

tacaccctgc cgccgcgtca ggtggctaac ggcgtagtgg acgcctttgt acacaccgtg 600

gaacagtatg ttaccaaacc ggttgatgcc aaaattcagg accgtttcgc agaaggcatt 660

ttgctgacgc taatcgaaga tggtccgaaa gccctgaaag agccagaaaa ctacgatgtg 720

cgcgccaacg tcatgtgggc ggcgactcag gcgctgaacg gtttgattgg cgctggcgta 780

ccgcaggact gggcaacgca tatgctgggc cacgaactga ctgcgatgca cggtctggat 840

cacgcgcaaa cactggctat cgtcctgcct gcactgtgga atgaaaaacg cgataccaag 900

cgcgctaagc tgctgcaata tgctgaacgc gtctggaaca tcactgaagg ttccgatgat 960

gagcgtattg acgccgcgat tgccgcaacc cgcaatttct ttgagcaatt aggcgtgccg 1020

acccacctct ccgactacgg tctggacggc agctccatcc cggctttgct gaaaaaactg 1080

gaagagcacg gcatgaccca actgggcgaa aatcatgaca ttacgttgga tgtcagccgc 1140

cgtatatacg aagccgcccg ctaa 1164

<210> 34

<211> 387

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 34

Met Asn Asn Phe Asn Leu His Thr Pro Thr Arg Ile Leu Phe Gly Lys

1 5 10 15

Gly Ala Ile Ala Gly Leu Arg Glu Gln Ile Pro His Asp Ala Arg Val

20 25 30

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

35 40 45

Gln Val Leu Asp Ala Leu Lys Gly Met Asp Val Leu Glu Phe Gly Gly

50 55 60

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

65 70 75 80

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

85 90 95

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

100 105 110

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

115 120 125

Ser Ala Ile Pro Met Gly Cys Val Leu Thr Leu Pro Ala Thr Gly Ser

130 135 140

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

145 150 155 160

Gln Ala Phe His Ser Ala His Val Gln Pro Val Phe Ala Val Leu Asp

165 170 175

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

180 185 190

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

195 200 205

Asp Ala Lys Ile Gln Asp Arg Phe Ala Glu Gly Ile Leu Leu Thr Leu

210 215 220

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

225 230 235 240

Arg Ala Asn Val Met Trp Ala Ala Thr Gln Ala Leu Asn Gly Leu Ile

245 250 255

Gly Ala Gly Val Pro Gln Asp Trp Ala Thr His Met Leu Gly His Glu

260 265 270

Leu Thr Ala Met His Gly Leu Asp His Ala Gln Thr Leu Ala Ile Val

275 280 285

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

290 295 300

Leu Gln Tyr Ala Glu Arg Val Trp Asn Ile Thr Glu Gly Ser Asp Asp

305 310 315 320

Glu Arg Ile Asp Ala Ala Ile Ala Ala Thr Arg Asn Phe Phe Glu Gln

325 330 335

Leu Gly Val Pro Thr His Leu Ser Asp Tyr Gly Leu Asp Gly Ser Ser

340 345 350

Ile Pro Ala Leu Leu Lys Lys Leu Glu Glu His Gly Met Thr Gln Leu

355 360 365

Gly Glu Asn His Asp Ile Thr Leu Asp Val Ser Arg Arg Ile Tyr Glu

370 375 380

Ala Ala Arg

385

<210> 35

<211> 828

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 35

atggctaatc caaccgttat taagctacag gatggcaatg tcatgcccca gctgggactg 60

ggcgtctggc aagcaagtaa tgaggaagta atcaccgcca ttcaaaaagc gttagaagtg 120

ggttatcgct cgattgatac cgccgcggcc tacaagaacg aagaaggtgt cggcaaagcc 180

ctgaaaaatg cctcagtcaa cagagaagaa ctgttcatca ccactaagct gtggaacgac 240

gaccacaagc gcccccgcga agccctgctc gacagcctga aaaaactcca gcttgattat 300

atcgacctct acttaatgca ctggcccgtt cccgctatcg accattatgt cgaagcatgg 360

aaaggcatga tcgaattgca aaaagaggga ttaatcaaaa gcatcggcgt gtgcaacttc 420

cagatccatc acctgcaacg cctgattgat gaaactggcg tgacgcctgt gataaaccag 480

atcgaacttc atccgctgat gcaacaacgc cagctacacg cctggaacgc gacacacaaa 540

atccagaccg aatcctggag cccattagcg caaggaggga aaggcgtttt cgatcagaaa 600

gtcattcgcg atctggcaga taaatacggc aaaaccccgg cgcagattgt tatccgctgg 660

catctggata gcggcctggt ggtgatcccg aaatcggtca caccttcacg tattgccgaa 720

aactttgatg tctgggattt ccgtctcgac aaagacgaac tcggcgaaat tgcaaaactc 780

gatcagggca agcgtctcgg tcccgatcct gaccagttcg gcggctaa 828

<210> 36

<211> 275

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 36

Met Ala Asn Pro Thr Val Ile Lys Leu Gln Asp Gly Asn Val Met Pro

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

Ser Val Asn Arg Glu Glu Leu Phe Ile Thr Thr Lys Leu Trp Asn Asp

65 70 75 80

Asp His Lys Arg Pro Arg Glu Ala Leu Leu Asp Ser Leu Lys Lys Leu

85 90 95

Gln Leu Asp Tyr Ile Asp Leu Tyr Leu Met His Trp Pro Val Pro Ala

100 105 110

Ile Asp His Tyr Val Glu Ala Trp Lys Gly Met Ile Glu Leu Gln Lys

115 120 125

Glu Gly Leu Ile Lys Ser Ile Gly Val Cys Asn Phe Gln Ile His His

130 135 140

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

145 150 155 160

Ile Glu Leu His Pro Leu Met Gln Gln Arg Gln Leu His Ala Trp Asn

165 170 175

Ala Thr His Lys Ile Gln Thr Glu Ser Trp Ser Pro Leu Ala Gln Gly

180 185 190

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

195 200 205

Tyr Gly Lys Thr Pro Ala Gln Ile Val Ile Arg Trp His Leu Asp Ser

210 215 220

Gly Leu Val Val Ile Pro Lys Ser Val Thr Pro Ser Arg Ile Ala Glu

225 230 235 240

Asn Phe Asp Val Trp Asp Phe Arg Leu Asp Lys Asp Glu Leu Gly Glu

245 250 255

Ile Ala Lys Leu Asp Gln Gly Lys Arg Leu Gly Pro Asp Pro Asp Gln

260 265 270

Phe Gly Gly

275

<210> 37

<211> 1050

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 37

atgaagatca aagctgttgg tgcatattcc gctaaacaac cacttgaacc gatggatatc 60

acccggcgtg aaccgggacc gaatgatgtc aaaatcgaaa tcgcttactg tggcgtttgc 120

cattccgatc tccaccaggt ccgttccgag tgggcgggga cggtttaccc ctgcgtgccg 180

ggtcatgaaa ttgtggggcg tgtggtagcc gttggtgatc aggtagaaaa atatgcgccg 240

ggcgatctgg tcggtgtcgg ctgcattgtc gacagttgta aacattgcga agagtgtgaa 300

gacgggttgg aaaactactg tgatcacatg accggcacct ataactcgcc gacgccggac 360

gaaccgggcc atactctggg cggctactca caacagatcg tcgttcatga gcgatatgtt 420

ctgcgtattc gtcacccgca agagcagctg gcggcggtgg ctcctttgtt gtgtgcaggg 480

atcaccacgt attcgccgct acgtcactgg caggccgggc cgggtaaaaa agtgggcgtg 540

gtcggcatcg gcggtctggg acatatgggg attaagctgg cccacgcgat gggggcacat 600

gtggtggcat ttaccacttc tgaggcaaaa cgcgaagcgg caaaagccct gggggccgat 660

gaagttgtta actcacgcaa tgccgatgag atggcggctc atctgaagag tttcgatttc 720

attttgaata cagtagctgc gccacataat ctcgacgatt ttaccacctt gctgaagcgt 780

gatggcacca tgacgctggt tggtgcgcct gcgacaccgc ataaatcgcc ggaagttttc 840

aacctgatca tgaaacgccg tgcgatagcc ggttctatga ttggcggcat tccagaaact 900

caggagatgc tcgatttttg cgccgaacat ggcatcgtgg ctgatataga gatgattcgg 960

gccgatcaaa ttaatgaagc ctatgagcga atgctgcgcg gtgatgtgaa atatcgtttt 1020

gttatcgata atcgcacact aacagactga 1050

<210> 38

<211> 349

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 38

Met Lys Ile Lys Ala Val Gly Ala Tyr Ser Ala Lys Gln Pro Leu Glu

1 5 10 15

Pro Met Asp Ile Thr Arg Arg Glu Pro Gly Pro Asn Asp Val Lys Ile

20 25 30

Glu Ile Ala Tyr Cys Gly Val Cys His Ser Asp Leu His Gln Val Arg

35 40 45

Ser Glu Trp Ala Gly Thr Val Tyr Pro Cys Val Pro Gly His Glu Ile

50 55 60

Val Gly Arg Val Val Ala Val Gly Asp Gln Val Glu Lys Tyr Ala Pro

65 70 75 80

Gly Asp Leu Val Gly Val Gly Cys Ile Val Asp Ser Cys Lys His Cys

85 90 95

Glu Glu Cys Glu Asp Gly Leu Glu Asn Tyr Cys Asp His Met Thr Gly

100 105 110

Thr Tyr Asn Ser Pro Thr Pro Asp Glu Pro Gly His Thr Leu Gly Gly

115 120 125

Tyr Ser Gln Gln Ile Val Val His Glu Arg Tyr Val Leu Arg Ile Arg

130 135 140

His Pro Gln Glu Gln Leu Ala Ala Val Ala Pro Leu Leu Cys Ala Gly

145 150 155 160

Ile Thr Thr Tyr Ser Pro Leu Arg His Trp Gln Ala Gly Pro Gly Lys

165 170 175

Lys Val Gly Val Val Gly Ile Gly Gly Leu Gly His Met Gly Ile Lys

180 185 190

Leu Ala His Ala Met Gly Ala His Val Val Ala Phe Thr Thr Ser Glu

195 200 205

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

210 215 220

Ser Arg Asn Ala Asp Glu Met Ala Ala His Leu Lys Ser Phe Asp Phe

225 230 235 240

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

245 250 255

Leu Leu Lys Arg Asp Gly Thr Met Thr Leu Val Gly Ala Pro Ala Thr

260 265 270

Pro His Lys Ser Pro Glu Val Phe Asn Leu Ile Met Lys Arg Arg Ala

275 280 285

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

290 295 300

Asp Phe Cys Ala Glu His Gly Ile Val Ala Asp Ile Glu Met Ile Arg

305 310 315 320

Ala Asp Gln Ile Asn Glu Ala Tyr Glu Arg Met Leu Arg Gly Asp Val

325 330 335

Lys Tyr Arg Phe Val Ile Asp Asn Arg Thr Leu Thr Asp

340 345

<210> 39

<211> 1020

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 39

atgtcgatga taaaaagcta tgccgcaaaa gaagcgggcg gcgaactgga agtttatgag 60

tacgatcccg gtgagctgag gccacaagat gttgaagtgc aggtggatta ctgcgggatc 120

tgccattccg atctgtcgat gatcgataac gaatggggat tttcacaata tccgctggtt 180

gccgggcatg aggtgattgg gcgcgtggtg gcactcggga gcgccgcgca ggataaaggt 240

ttgcaggtcg gtcagcgtgt cgggattggc tggacggcgc gtagctgtgg tcactgcgac 300

gcctgtatta gcggtaatca gatcaactgc gagcaaggtg cggtgccgac gattatgaat 360

cgcggtggct ttgccgagaa gttgcgtgcg gactggcaat gggtgattcc actgccagaa 420

aatattgata tcgagtccgc cgggccgctg ttgtgcggcg gtatcacggt ctttaaacca 480

ctgttgatgc accatatcac tgctaccagc cgcgttgggg taattggtat tggcgggctg 540

gggcatatcg ctataaaact tctgcacgca atgggatgcg aggtgacagc ctttagttct 600

aatccggcga aagagcagga agtgctggcg atgggtgccg ataaagtggt gaatagccgc 660

gatccgcagg cactgaaagc actggcgggg cagtttgatc tcattatcaa caccgtcaac 720

gtcagcctcg actggcagcc ctattttgag gcgctgacct atggcggtaa tttccatacg 780

gtcggtgcgg ttctcacgcc gctgtctgtt ccggccttta cgttaattgc gggcgatcgc 840

agcgtctctg gttctgctac cggcacgcct tatgagctgc gtaagctgat gcgttttgcc 900

gcccgcagca aggttgcgcc gaccaccgaa ctgttcccga tgtcgaaaat taacgacgcc 960

atccagcatg tgcgcgacgg taaggcgcgt taccgcgtgg tgttgaaagc cgatttttga 1020

<210> 40

<211> 339

<212> PRT

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 40

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

1 5 10 15

Glu Val Tyr Glu Tyr Asp Pro Gly Glu Leu Arg Pro Gln Asp Val Glu

20 25 30

Val Gln Val Asp Tyr Cys Gly Ile Cys His Ser Asp Leu Ser Met Ile

35 40 45

Asp Asn Glu Trp Gly Phe Ser Gln Tyr Pro Leu Val Ala Gly His Glu

50 55 60

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

65 70 75 80

Leu Gln Val Gly Gln Arg Val Gly Ile Gly Trp Thr Ala Arg Ser Cys

85 90 95

Gly His Cys Asp Ala Cys Ile Ser Gly Asn Gln Ile Asn Cys Glu Gln

100 105 110

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

115 120 125

Arg Ala Asp Trp Gln Trp Val Ile Pro Leu Pro Glu Asn Ile Asp Ile

130 135 140

Glu Ser Ala Gly Pro Leu Leu Cys Gly Gly Ile Thr Val Phe Lys Pro

145 150 155 160

Leu Leu Met His His Ile Thr Ala Thr Ser Arg Val Gly Val Ile Gly

165 170 175

Ile Gly Gly Leu Gly His Ile Ala Ile Lys Leu Leu His Ala Met Gly

180 185 190

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

195 200 205

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

210 215 220

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

225 230 235 240

Val Ser Leu Asp Trp Gln Pro Tyr Phe Glu Ala Leu Thr Tyr Gly Gly

245 250 255

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

260 265 270

Phe Thr Leu Ile Ala Gly Asp Arg Ser Val Ser Gly Ser Ala Thr Gly

275 280 285

Thr Pro Tyr Glu Leu Arg Lys Leu Met Arg Phe Ala Ala Arg Ser Lys

290 295 300

Val Ala Pro Thr Thr Glu Leu Phe Pro Met Ser Lys Ile Asn Asp Ala

305 310 315 320

Ile Gln His Val Arg Asp Gly Lys Ala Arg Tyr Arg Val Val Leu Lys

325 330 335

Ala Asp Phe

<210> 41

<211> 62

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 41

atggcccaac aatcacccta ttcagcagcg atggcagaac aggtgtaggc tggagctgct 60

tc 62

<210> 42

<211> 60

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 42

tcaatcgctt ttcagcaaca cctcttccag ccactggcgc acatatgaat atcctcctta 60

<210> 43

<211> 60

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 43

atggctatcc ctgcatttgg tttaggtact ttccgtctga gtgtaggctg gagctgcttc 60

<210> 44

<211> 62

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 44

ttaatcccat tcaggagcca gaccttccgg gctaaccagg cggcatatga atatcctcct 60

ta 62

<210> 45

<211> 62

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 45

atgcaacaaa aaatgattca atttagtggc gatgtctcac tggtgtaggc tggagctgct 60

tc 62

<210> 46

<211> 64

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 46

atgctacaaa attgcgcaca atcaaattgc cgcattattc ctaagtgtag gctggagctg 60

cttc 64

<210> 47

<211> 61

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 47

ttaattcccc tgcatcgccc gcattcttgc cgcatcttcc cccatatgaa tatcctcctt 60

a 61

<210> 48

<211> 61

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 48

atgaacaact ttaatctgca caccccaacc cgcattctgt tgtgtaggct ggagctgctt 60

c 61

<210> 49

<211> 60

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 49

ttagcgggcg gcttcgtata tacggcggct gacatccaac gcatatgaat atcctcctta 60

<210> 50

<211> 62

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 50

atggctaatc caaccgttat taagctacag gatggcaatg tcgtgtaggc tggagctgct 60

tc 62

<210> 51

<211> 60

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 51

ttagccgccg aactggtcag gatcgggacc gagacgcttg ccatatgaat atcctcctta 60

<210> 52

<211> 61

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 52

atgaagatca aagctgttgg tgcatattcc gctaaacaac cgtgtaggct ggagctgctt 60

c 61

<210> 53

<211> 60

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 53

tcagtctgtt agtgtgcgat tatcgataac aaaacgatat tcatatgaat atcctcctta 60

<210> 54

<211> 62

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 54

atgtcgatga taaaaagcta tgccgcaaaa gaagcgggcg gcgtgtaggc tggagctgct 60

tc 62

<210> 55

<211> 63

<212> DNA

<213> 2 Ambystoma laterale x Ambystoma jeffersonianum

<400> 55

tcaaaaatcg gctttcaaca ccacgcggta acgcgcctta ccgtcatatg aatatcctcc 60

tta 63

Claims (5)

1. a kind of method of microbial fermentation glucose synthesis phenylacetic acid, p-hydroxyphenylaceticacid and heteroauxin, it is characterised in that:

Knocked out in microorganism TrpR gene or its function equivalents, dkgB gene or its function equivalents, yeaE gene or its Function equivalents, yqhC gene or its function equivalents, yqhD gene or its function equivalents, dkgA gene or its function are equivalent Body, yahK gene or its function equivalents and yjgB gene or its function equivalents, by KDC gene or its function equivalents, ALDH gene or its function equivalents and ARO8 gene or its function equivalents, aroG^fbrGene or its function equivalents and pheA^fbrThe carrier is transferred in gene knock-out bacterial strain to carrier, thus obtains target by gene or the recombination of its function equivalents Microorganism can realize that glucose fermentation synthesizes phenylacetic acid；

Or replacement pheA^fbrGene or its function equivalents are tyrA^fbrGene or its function equivalents, the target thus modified are micro- The achievable glucose fermentation of biology synthesizes p-hydroxyphenylaceticacid；

Or replacement pheA^fbrGene or its function equivalents are TrpE^fbrGene or its function equivalents, serA^fbrGene or its function Energy equivalents and trpABCD gene or its function equivalents, the objective microbe thus modified can realize that glucose fermentation synthesizes Heteroauxin.

2. a kind of method of synthesis phenylacetic acid in biology, produces phenylacetic acid by raw material of glucose, it is characterised in that want using right Microorganism described in asking 1 converts glucose into phenylacetic acid.

3. a kind of method of synthesis p-hydroxyphenylaceticacid in biology, produces p-hydroxyphenylaceticacid by raw material of glucose, feature exists P-hydroxyphenylaceticacid is converted glucose into use microorganism described in claim 1.

4. the method for synthesis of indole acetic acid, produces by raw material of glucose to heteroauxin in a kind of biology, it is characterised in that use Microorganism described in claim 1 converts glucose into phenylacetic acid.

5. microorganism according to claim 1, it is characterised in that: the microorganism includes the nucleic acid sequence of ALDH gene； The nucleic acid sequence of the coding ALDH gene can be catalyzed phenylacetaldehyde, para hydroxybenzene acetaldehyde and heteroauxin and be separately converted to benzene second Acid, p-hydroxyphenylaceticacid and heteroauxin.