CN1928085A - Gene sequence of penaeus monodon cell periodic protein B - Google Patents
Gene sequence of penaeus monodon cell periodic protein B Download PDFInfo
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- CN1928085A CN1928085A CN 200610037559 CN200610037559A CN1928085A CN 1928085 A CN1928085 A CN 1928085A CN 200610037559 CN200610037559 CN 200610037559 CN 200610037559 A CN200610037559 A CN 200610037559A CN 1928085 A CN1928085 A CN 1928085A
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Abstract
The present invention designs degenerating primer in the conservation region of CDS sequence in kindred species cyclin B gene to extract total RNA from gonad of tiger prawn, and clones to the expression sequence of cyclin B gene of prawn through PCR process amplification. By means of cloning the cyclin B gene, it is possible to understand the gene composition and the growth and development cycle process of fish. Through obtaining the gene purified protein, it is possible to promote fish cell growth, shorten fish growth period and raise economic benefit.
Description
Technical field
The present invention relates to a kind of tigar prawn cell periodic protein B gene order.
Background technology
Cell cycle is meant that cycling cell finishes the whole process that is experienced to the end of mitotic division next time from a mitotic division.In this process, the cytogenetics material duplicates and doubles, and when division finishes evenly distribute in two daughter cells.Cell cycle can be divided into interval and m period again.Since a mitotic division finish to the period of mitotic division next time be exactly interval.This first phase synthesizes sign with DNA, and through division stage, the karyomit(e) that doubles and other cellular components are by in evenly distribute to the two duplicate daughter cell.By division, formed a new cell.Cell cycle can be inseparable with the orderly expression of relevant regulatory gene to the sequential operation of m period in strict accordance with interval.This gene can induce immature cell to enter m period, and the unique synthetic gene protein that needs of concurrent present m period is exactly this cyclin, and this is a fissional adjusting albumen that plays a crucial role.
Cell periodic protein B is the m period cyclin, brings into play function in interval to the m period intersection, the inducing cell division.Cyclin and cell aging are in close relations, and the cell periodic protein B degraded is obstructed, and cell can not withdraw from m period, influences cell aging.
Summary of the invention
The objective of the invention is by degenerate primer, and with the amplification of PCR method, be cloned into the full expressed sequence of the gene of tigar prawn cell periodic protein B in conjunction with the RACE technology with the conserved regions design of the CDS sequence of the gene of nearly source species cell periodic protein B.
Above-mentioned purpose is achieved through the following technical solutions:
Dissect prawn and take out sexual gland, extract the total RNA of tigar prawn, make it to mix, carry out reverse transcription, the synthetic first chain cDNA with reverse transcription primer (oligo-dT joint primer).
From GenBank, download nearly source species (as people, ox, zebra fish, japonicus etc.) cell periodic protein B homologous gene CDS sequence, utilize Clustal W software to carry out the multisequencing comparison, determine conserved regions, according to conserved regions sequences Design degenerate primer, the amplified fragments size is about 225bp.Adopt β-second eyeball phosphoramidite chemical method to carry out that DNA is synthetic to obtain following primer sequence after the design of primers:
F：5’ATWGCWAGYAAATAYGAAGARATGTA?3’
R：5’TCCATIARRTAYTTKGCWARIGTATG?3’。
As template, carry out pcr amplification with degenerate primer with the synthetic first chain cDNA, institute's amplification PCR products detects with 1.2% agarose gel electrophoresis, and purifying reclaims the purpose product from gel.Then with the PCR product cloning of purifying in the pMD-18T carrier, transformed into escherichia coli JM-109 competent cell, the picking positive colony extracts plasmid DNA.After degenerated primer PCR detects, will have the segmental plasmid DNA of insertion and carry out two-way order-checking with the M13 universal primer.
According to the first chain cDNA fragment sequence that obtained design Auele Specific Primer, utilize the terminal rapid amplifying technology of cDNA (Rapid Amplification of cDNA ends, RACE) to 3 of goal gene ' and 5 ' end carry out pcr amplification.
In the rapid amplifying of 3 ' end, carry out the PCR reaction by gene primer and joint primer, the amplified fragments size is about 1000bp.Described gene primer and joint primer sequence are
Gene primer: 5 ' TCGGGGACTTCGCATACATC 3 '
Joint primer: 5 ' GGCCACGCGACTAGTAC 3 '.
In the rapid amplifying of 5 ' end, utilize terminal enzyme (DNA) and dCTP after the cDNA end adds poly (C) tail, with the cDNA behind the tailing as template, utilize outside Auele Specific Primer and OligodG to carry out the pcr amplification first time, gained PCR product utilizes inboard primer and OligodG to carry out the pcr amplification second time again.Described primer sequence is:
oligo-dG：5’GGGGGGGGGGGGGGGH?3’
Gene outside primer: 5 ' GCAAAGGTATGTTGAGAAGC 3 '
The inboard primer of gene: 5 ' GTGTAAGGGAAGGGGATAGG 3 '.
After terminal rapid amplifying technology carries out carrying out separation detection on the agarose gel electrophoresis of the resulting PCR product of PCR 1.2%, behind the PCR product purification, be cloned in the pMD-18T carrier, transformed into escherichia coli JM-109 competent cell, the picking positive colony, with the M13 primer plasmid DNA is checked order, order-checking institute calling sequence utilizes Clustal W software to splice with the sequence of degenerate primer amplification gained again can obtain goal gene.
Clone by cell cycle protein B gene, understand the composition situation and fish growth process growth cycle of gene, and, can promote the fish somatocyte development by to the proteic acquisition of this isogeneity, shorten fish growth cycles, to certain promoter action of increasing economic efficiency.
Embodiment
The present invention is further elaborated below by following embodiment, but content of the present invention is not limited to this fully.
1. the extraction of total RNA
Get fresh and alive healthy tigar prawn (the about 300g of body weight) and in the laboratory, support temporarily (about 24 ℃ of water temperature behind the 2d, air-pump inflating), dissects the shrimp body and take out the about 100mg of sexual gland, put into 1mL Trizol (Gibco, Japan) carry out homogenate in, extract total RNA according to the test kit operation instruction.
2.cDNA first chain is synthetic
Getting the total RNA 5 μ g of tigar prawn mixes with reverse transcription primer (oligo-dT joint primer) 1 μ L (10pmol/L), behind 70 ℃ of heating 5min, place on ice immediately, add 5 * buffer then, 2.5mmol/L dNTP mixed solution, Ribonuclease Inhibitor, M-MLV ThermoScript II, reaction system are 25 μ L.Reaction process is 42 ℃ of 60min, 70 ℃ of 15min, and it is standby to put into-80 ℃ of preservations at last.
3. degenerate primer design considerations, primer synthetic method
From GenBank, download nearly source species (as people, ox, zebra fish, japonicus etc.) cell periodic protein B homologous gene CDS sequence, utilize Clustal W software to carry out the multisequencing comparison, determine conserved regions, according to conserved regions sequences Design degenerate primer, the amplified fragments size is about 225bp.Adopt β-second eyeball phosphoramidite chemical method to carry out DNA after the design of primers and synthesize, obtain following primer sequence:
F：5’ATWGCWAGYAAATAYGAAGARATGTA?3’
R；5’TCCATIARRTAYTTKGCWARIGTATG?3’。
4. the clone of cell periodic protein B gene cDNA complete sequence
4.1 the first chain cDNA carries out pcr amplification with degenerate primer
With the degenerate primer of the conserved regions design of nearly source species cell periodic protein B gene C DS sequence, the amplified fragments size is about 225bp.
As template, carry out pcr amplification with degenerate primer with the above-mentioned synthetic first chain cDNA, reaction system is: 10x PCR reaction buffer 5 μ L, 25mmol/L MgCl
23 μ L, 2.5mmol/L dNTP 2 μ L, each 2 μ L of 10nmol/L primer dTF and dTR, Taq enzyme 1.25U is supplemented to 50 μ L with PCR water with reaction system.Reaction conditions is: 1 circulation, 94 ℃ of sex change 5min; 35 circulations: 94 ℃ of sex change 45s, 56 ℃ of annealing 45s, 72 ℃ are extended 45s; 1 circulation, 72 ℃ are extended 10min; 4 ℃ of insulations.Institute's amplification PCR products detects with 1.2% agarose gel electrophoresis, and purifying reclaims the purpose product from gel.Then with the PCR product cloning of purifying in the pMD-18T carrier, transformed into escherichia coli JM-109 competent cell, the picking positive colony extracts plasmid DNA.After degenerated primer PCR detects, will have the segmental plasmid DNA of insertion and carry out two-way order-checking with the M13 universal primer.
4.2 the rapid amplifying technology of the first chain cDNA is carried out pcr amplification
According to the cDNA fragment sequence design Auele Specific Primer that has obtained.Utilize the terminal rapid amplifying technology of cDNA (Rapid Amplification of cDNA ends, RACE) to 3 of goal gene ' and 5 ' end carry out pcr amplification.
In 3 ' RACE, carry out pcr amplification by gene primer and joint primer.
Primer sequence is:
Gene primer: 5 ' TCGGGGACTTCGCATACATC 3 '
Joint primer: 5 ' GGCCACGCGACTAGTAC 3 '.
Reaction system is with the degenerate primer amplification system.
Reaction conditions: 1 circulation, 94 ℃ of sex change 5min; 35 circulations: 94 ℃ of sex change 45s, 58 ℃ of annealing 30s, 72 ℃ are extended 1min; 1 circulation, 72 ℃ are extended 10min; 4 ℃ of insulations.
In 5 ' RACE, utilize terminal enzyme (DNA) and dCTP after the cDNA end adds poly (C) tail, with the cDNA behind the tailing as template, utilize outside Auele Specific Primer and OligodG to carry out the pcr amplification first time, gained PCR product is got 1 μ L as template, utilizes inboard primer and OligodG to carry out the pcr amplification second time again.
Primer sequence is:
oligo-dG：5’GGGGGGGGGGGGGGGH?3’
Gene outside primer: 5 ' GCAAAGGTATGTTGAGAAGC 3 '
The inboard primer of gene: 5 ' GTGTAAGGGAAGGGGATAGG 3 '.
Reaction system is with the degenerate primer amplification system.
Reaction conditions:
PCR reaction for the first time: 1 circulation, 94 ℃ of sex change 5min; 35 circulations: 94 ℃ of sex change 45s, 58 ℃ of annealing 30s, 72 ℃ are extended 1min; 1 circulation, 72 ℃ are extended 10min; 4 ℃ of insulations.
PCR reaction for the second time: 1 circulation, 94 ℃ of sex change 5min; 35 circulations: 94 ℃ of sex change 45s, 59 ℃ of annealing 30s, 72 ℃ are extended 1min; 1 circulation, 72 ℃ are extended 10min; 4 ℃ of insulations.
Resulting PCR product is after carrying out separation detection on 1.2% the agarose gel electrophoresis, behind the PCR product purification, be cloned in the pMD-18T carrier, transformed into escherichia coli JM-109 competent cell, the picking positive colony, with the M13 primer plasmid DNA is checked order, order-checking institute calling sequence utilizes Clustal W software to splice with the sequence of degenerate primer amplification gained again can obtain goal gene.
5. to the mensuration of tigar prawn cell periodic protein B gene
Resulting PCR product is after carrying out separation detection on 1.2% the agarose gel electrophoresis, behind the PCR product purification, be cloned in the pMD-18T carrier transformed into escherichia coli JM-109 competent cell, the picking positive colony checks order to plasmid DNA with 3730 sequenators.Sequencing result BLAST software (
Http:// www.ncbi.nim.nih.gov/) carry out homology mensuration, be defined as tigar prawn cell periodic protein B dna homolog sequence.Comparison result is:
Score E
Sequences?producing?significant?alignments (Bits) Value
gi|54660743|gb|AAV37462.1|cyclin?B[Marsupenaeus?japonicus]
528 1e-148
gi|10955|emb|CAA41255.1|cyclin?B[Patella?vulgata]＞gi|11617...
319 9e-86
gi|52851368|dbj|BAD52077.1|cyclin?B2[Anguilla?japonica]＞gi...
305 2e-81
gi|10337|emb|CAA33513.1|unnamed?protein?product[Spisula?sol...
303 9e-81
gi|42542462|gb|AAH66507.1|Cyclin?B2[Danio?rerio]＞gi|282778...
302 1e-80
gi|68363504|ref|XP_709645.1|PREDICTED：similar?to?Cyclin?B2?iso
301 3e-80
According to the report of Iron1995,, prove that promptly sequence is with a kind of gene order as long as then have absolute homology between two sequences with E-Value value after the comparison of BLAST software less than 0.005 between two sequences.After seeing tigar prawn target gene sequences BLAST comparison from comparing result, with the E value of the cell periodic protein B of other species definitely less than 0.005, so prove that the goal gene of being cloned is the homologous sequence of cell periodic protein B.
Sequence table
＜110〉Nanhai Aquatic Inst., Chinese Aquatic Scientific Research Inst
＜120〉gene order of tigar prawn cell periodic protein B
<160>2
<210>1
<211>1658
<212>RNA
＜213〉tigar prawn (Penaeus monodon Fabricius)
<220>
<221>3’UTP
<222>(1322)...(1658)
<220>
<221>5’UTP
<222>(1)...(59)
<220>
<221>CDS
<222>(60)...(1262)
<220>
<221>PolyA?site
<222>(1641)...(1658)
<220>
<221>PolyA?signal
<222>(1642)...(1658)
<400>1
ggagatttca?gagttaataa?agcctcgcat?cgttgattac?cggttgattt?ccgttgatc 59
atg?tgt?tta?aga?act?acc?tcg?cat?ctt?agt?aat?gtg?gga?cat?gag?ctg 107
Met?Cys?Leu?Arg?Thr?Thr?Ser?His?Leu?Ser?Asn?Val?Gly?His?Glu?Leu
1 7 13
aac?aac?cca?cgc?aaa?gta?gag?gca?aag?atg?atc?cag?ggg?cca?gtc?gcc 155
Asn?Asn?Pro?Arg?Lys?Val?Glu?Ala?Lys?Met?Ile?Gln?Gly?Pro?Val?Ala
17 23 29
cgt?cgt?gcc?ttg?ggg?gat?gtt?ggc?aac?cat?ggt?att?ccc?gtg?caa?ggg 203
Arg?Arg?Ala?Leu?Gly?Asp?Val?Gly?Asn?His?Gly?Ile?Pro?Val?Gln?Gly
33 39 45
ccc?aaa?gct?cct?ctc?aag?gcg?ggg?gag?atc?tct?cgc?aat?gag?ccc?gtc 251
Pro?Lys?Ala?Pro?Leu?Lys?Ala?Gly?Glu?Ile?Ser?Arg?Asn?Glu?Pro?Val
49 55 61
aag?ctg?cac?aag?ccc?aag?tcc?ggc?ctc?tct?ggg?ctg?ctc?gcc?aga?ccc 299
Lys?Leu?His?Lys?Pro?Lys?Ser?Gly?Leu?Ser?Gly?Leu?Leu?Ala?Arg?Pro
65 71 77
ggc?aaa?gaa?aat?gtg?aag?ccc?ctg?aag?gaa?gtg?gta?gag?cat?gtg?gag 347
Gly?Lys?Glu?Asn?Val?Lys?Pro?Leu?Lys?Glu?Val?Val?Glu?His?Val?Glu
81 87 93
cag?atg?gat?gtg?gag?gag?gaa?gcc?aaa?gtg?gaa?gag?ctg?gct?att?gct 395
Gln?Met?Asp?Val?Glu?Glu?Glu?Ala?Lys?Val?Glu?Glu?Leu?Ala?Ile?Ala
97 103 109
ttc?tct?acc?cag?aga?cta?gat?gtt?gaa?gat?att?gat?gcc?caa?gac?agt 443
Phe?Ser?Thr?Gln?Arg?Leu?Asp?Val?Glu?Asp?Ile?Asp?Ala?Gln?Asp?Ser
113 119 125
gat?aat?cct?cag?ctt?gta?tct?gaa?tat?gtg?aat?gat?atc?tac?aag?tac 491
Asp?Asn?Pro?Gln?Leu?Val?Ser?Glu?Tyr?Val?Asn?Asp?Ile?Tyr?Lys?Tyr
129 135 141
ctg?cga?gag?ctg?gag?gat?gcc?aac?aaa?gtc?aag?ccc?aga?tac?cta?gaa 539
Leu?Arg?Glu?Leu?Glu?Asp?Ala?Asn?Lys?Val?Lys?Pro?Arg?Tyr?Leu?Glu
145 151 157
ggc?caa?gta?att?aca?gga?aag?atg?aga?gca?att?ttg?att?gac?tgg?ctt 587
Gly?Gln?Val?Ile?Thr?Gly?Lys?Met?Arg?Ala?Ile?Leu?Ile?Asp?Trp?Leu
161 167 173
gtc?caa?gta?cac?ctc?cgc?ttc?act?ctg?ctt?caa?gaa?aca?ctg?tat?ctg 635
Val?Gln?Val?His?Leu?Arg?Phe?Thr?Leu?Leu?Gln?Glu?Thr?Leu?Tyr?Leu
177 183 189
act?gtt?gct?atc?att?gac?aga?ttt?ctc?cag?act?cag?agg?aat?ata?cca 683
Thr?Val?Ala?Ile?Ile?Asp?Arg?Phe?Leu?Gln?Thr?Gln?Arg?Asn?Ile?Pro
193 199 205
cgt?aac?aag?cta?cag?tta?gtt?ggt?gtg?act?gcc?atg?ttc?ata?gct?agt 731
Arg?Asn?Lys?Leu?Gln?Leu?Val?Gly?Val?Thr?Ala?Met?Phe?Ile?Ala?Ser
209 215 221
aaa?tat?gaa?gaa?atg?tat?tgc?cca?gaa?atc?ggg?gac?ttc?gca?tac?atc 779
Lys?Tyr?Glu?Glu?Met?Tyr?Cys?Pro?Glu?Ile?Gly?Asp?Phe?Ala?Tyr?Ile
225 231 237
aca?gac?aaa?gcc?tac?tca?aag?gca?gag?att?cgt?aaa?atg?gag?gtg?acc 827
Thr?Asp?Lys?Ala?Tyr?Ser?Lys?Ala?Glu?Ile?Arg?Lys?Met?Glu?Val?Thr
241 247 253
atg?ctg?aat?gag?ctg?ggc?ttc?aat?gta?tcc?tat?ccc?ctt?ccc?tta?cac 875
Met?Leu?Asn?Glu?Leu?Gly?Phe?Asn?Val?Ser?Tyr?Pro?Leu?Pro?Leu?His
257 263 269
ttc?ctg?cga?aga?aac?agc?aaa?gct?ggc?tct?gtt?gat?gct?tct?caa?cac 923
Phe?Leu?Arg?Arg?Asn?Ser?Lys?Ala?Gly?Ser?Val?Asp?Ala?Ser?Gln?His
273 279 285
acc?ttg?gca?aag?tac?ttg?atg?gaa?ctt?tgc?ttg?cca?gag?tac?agc?atg 971
Thr?Leu?Ala?Lys?Tyr?Leu?Met?Glu?Leu?Cys?Leu?Pro?Glu?Tyr?Ser?Met
289 295 301
tgc?cat?tac?aag?tca?tca?atg?att?gct?gca?tct?gct?ctc?tgc?ctt?tca 1019
Cys?His?Tyr?Lys?Ser?Ser?Met?Ile?Ala?Ala?Ser?Ala?Leu?Cys?Leu?Ser
305 311 317
ctt?aaa?ttg?ttg?gat?ggc?aat?aac?tgg?agt?gat?aca?ttg?act?ttc?tat 1067
Leu?Lys?Leu?Leu?Asp?Gly?Asn?Asn?Trp?Ser?Asp?Thr?Leu?Thr?Phe?Tyr
321 327 333
tct?cgc?tac?act?gaa?caa?cag?ctc?atg?cca?gtc?atg?tgc?aaa?atg?gca 1115
Ser?Arg?Tyr?Thr?Glu?Gln?Gln?Leu?Met?Pro?Val?Met?Cys?Lys?Met?Ala
337 343 349
tca?gtt?gta?gta?aag?agc?agt?agt?gcc?aag?caa?cag?gct?gta?aga?cag 1163
Ser?Val?Val?Val?Lys?Ser?Ser?Ser?Ala?Lys?Gln?Gln?Ala?Val?Arg?Gln
353 359 365
aag?tac?aaa?gcc?agc?aag?ttg?atg?aaa?att?agt?gag?att?ccc?cag?ctc 1211
Lys?Tyr?Lys?Ala?Ser?Lys?Leu?Met?Lys?Ile?Ser?Glu?Ile?Pro?Gln?Leu
369 375 381
aaa?tcg?aag?ctc?atc?aac?tcg?ctc?gca?gag?aag?agt?gcg?tct?tat?gca 1259
Lys?Ser?Lys?Leu?Ile?Asn?Ser?Leu?Ala?Glu?Lys?Ser?Ala?Ser?Tyr?Ala
385 391 397
tga 1262
*
ggtggtcgcc?atttaaaaag?taaatattgt?tcatgttgaa?agaatggggt?tgtttttgta 1322
catatttttt?taagacgcca?gttttttttt?ttcataagtt?ttcagattta?taaaatactg 1382
accaccagtt?tttttttttt?tttttaacaa?gagtagtttt?ggggggcagc?atgtaccccc 1442
cagagtgggt?ggtttcggca?tccctccatt?catcgtggtc?ttcattgtat?taaaataatt 1502
ttaggcagga?caaaaatggt?tatttgaaga?aaggaaccag?caaagaagta?tatgcaatgt 1562
cttgtaatgt?atatatgacc?tgtgataaac?ttttagttat?gttctgtatg?tatacctata 1622
taataaacca?gttttatcca?aaaaaaaaaa?aaaaaa 1658
<210>2
<211>400
<212>PRT
＜213〉tigar prawn (Penaeus monodon Fabricius)
<400>2
Met?Cys?Leu?Arg?Thr?Thr?Ser?His?Leu?Ser?Asn?Val?Gly?His?Glu?Leu
1 7 13
Asn?Asn?Pro?Arg?Lys?Val?Glu?Ala?Lys?Met?Ile?Gln?Gly?Pro?Val?Ala
17 23 29
Arg?Arg?Ala?Leu?Gly?Asp?Val?Gly?Asn?His?Gly?Ile?Pro?Val?Gln?Gly
33 39 45
Pro?Lys?Ala?Pro?Leu?Lys?Ala?Gly?Glu?Ile?Ser?Arg?Asn?Glu?Pro?Val
49 55 61
Lys?Leu?His?Lys?Pro?Lys?Ser?Gly?Leu?Ser?Gly?Leu?Leu?Ala?Arg?Pro
65 71 77
Gly?Lys?Glu?Asn?Val?Lys?Pro?Leu?Lys?Glu?Val?Val?Glu?His?Val?Glu
81 87 93
Gln?Met?Asp?Val?Glu?Glu?Glu?Ala?Lys?Val?Glu?Glu?Leu?Ala?Ile?Ala
97 103 109
Phe?Ser?Thr?Gln?Arg?Leu?Asp?Val?Glu?Asp?Ile?Asp?Ala?Gln?Asp?Ser
113 119 125
Asp?Asn?Pro?Gln?Leu?Val?Ser?Glu?Tyr?Val?Asn?Asp?Ile?Tyr?Lys?Tyr
129 135 141
Leu?Arg?Glu?Leu?Glu?Asp?Ala?Asn?Lys?Val?Lys?Pro?Arg?Tyr?Leu?Glu
145 151 157
Gly?Gln?Val?Ile?Thr?Gly?Lys?Met?Arg?Ala?Ile?Leu?Ile?Asp?Trp?Leu
161 167 173
Val?Gln?Val?His?Leu?Arg?Phe?Thr?Leu?Leu?Gln?Glu?Thr?Leu?Tyr?Leu
177 183 189
Thr?Val?Ala?Ile?Ile?Asp?Arg?Phe?Leu?Gln?Thr?Gln?Arg?Asn?Ile?Pro
193 199 205
Arg?Asn?Lys?Leu?Gln?Leu?Val?Gly?Val?Thr?Ala?Met?Phe?Ile?Ala?Ser
209 215 221
Lys?Tyr?Glu?Glu?Met?Tyr?Cys?Pro?Glu?Ile?Gly?Asp?Phe?Ala?Tyr?Ile
225 231 237
Thr?Asp?Lys?Ala?Tyr?Ser?Lys?Ala?Glu?Ile?Arg?Lys?Met?Glu?Val?Thr
241 247 253
Met?Leu?Asn?Glu?Leu?Gly?Phe?Asn?Val?Ser?Tyr?Pro?Leu?Pro?Leu?His
257 263 269
Phe?Leu?Arg?Arg?Asn?Ser?Lys?Ala?Gly?Ser?Val?Asp?Ala?Ser?Gln?His
273 279 285
Thr?Leu?Ala?Lys?Tyr?Leu?Met?Glu?Leu?Cys?Leu?Pro?Glu?Tyr?Ser?Met
289 295 301
Cys?His?Tyr?Lys?Ser?Ser?Met?Ile?Ala?Ala?Ser?Ala?Leu?Cys?Leu?Ser
305 311 317
Leu?Lys?Leu?Leu?Asp?Gly?Asn?Asn?Trp?Ser?Asp?Thr?Leu?Thr?Phe?Tyr
321 327 333
Ser?Arg?Tyr?Thr?Glu?Gln?Gln?Leu?Met?Pro?Val?Met?Cys?Lys?Met?Ala
337 343 349
Ser?Val?Val?Val?Lys?Ser?Ser?Ser?Ala?Lys?Gln?Gln?Ala?Val?Arg?Gln
353 359 365
Lys?Tyr?Lys?Ala?Ser?Lys?Leu?Met?Lys?Ile?Ser?Glu?Ile?Pro?Gln?Leu
369 375 381
Lys?Ser?Lys?Leu?Ile?Asn?Ser?Leu?Ala?Glu?Lys?Ser?Ala?Ser?Tyr?Ala
385 391 397
Claims (1)
1, a kind of tigar prawn cell periodic protein B gene is characterized in that having following RNA Nucleotide and amino acid sequence corresponding.
ggagatttca?gagttaataa?agcctcgcat?cgttgattac?cggttgattt?ccgttgatc 59
atg?tgt?tta?aga?act?acc?tcg?cat?ctt?agt?aat?gtg?gga?cat?gag?ctg 107
Met?Cys?Leu?Arg?Thr?Thr?Ser?His?Leu?Ser?Asn?Val?Gly?His?Glu?Leu
1 7 13
aac?aac?cca?cgc?aaa?gta?gag?gca?aag?atg?atc?cag?ggg?cca?gtc?gcc 155
Asn?Asn?Pro?Arg?Lys?Val?Glu?Ala?Lys?Met?Ile?Gln?Gly?Pro?Val?Ala
17 23 29
cgt?cgt?gcc?ttg?ggg?gat?gtt?ggc?aac?cat?ggt?att?ccc?gtg?caa?ggg 203
Arg?Arg?Ala?Leu?Gly?Asp?Val?Gly?Asn?His?Gly?Ile?Pro?Val?Gln?Gly
33 39 45
ccc?aaa?gct?cct?ctc?aag?gcg?ggg?gag?atc?tct?cgc?aat?gag?ccc?gtc 251
Pro?Lys?Ala?Pro?Leu?Lys?Ala?Gly?Glu?Ile?Ser?Arg?Asn?Glu?Pro?Val
49 55 61
aag?ctg?cac?aag?ccc?aag?tcc?ggc?ctc?tct?ggg?ctg?ctc?gcc?aga?ccc 299
Lys?Leu?His?Lys?Pro?Lys?Ser?Gly?Leu?Ser?Gly?Leu?Leu?Ala?Arg?Pro
65 71 77
ggc?aaa?gaa?aat?gtg?aag?ccc?ctg?aag?gaa?gtg?gta?gag?cat?gtg?gag 347
Gly?Lys?Glu?Asn?Val?Lys?Pro?Leu?Lys?Glu?Val?Val?Glu?His?Val?Glu
81 87 93
cag?atg?gat?gtg?gag?gag?gaa?gcc?aaa?gtg?gaa?gag?ctg?gct?att?gct 395
Gln?Met?Asp?Val?Glu?Glu?Glu?Ala?Lys?Val?Glu?Glu?Leu?Ala?Ile?Ala
97 103 109
ttc?tct?acc?cag?aga?cta?gat?gtt?gaa?gat?att?gat?gcc?caa?gac?agt 443
Phe?Ser?Thr?Gln?Arg?Leu?Asp?Val?Glu?Asp?Ile?Asp?Ala?Gln?Asp?Ser
113 119 125
gat?aat?cct?cag?ctt?gta?tct?gaa?tat?gtg?aat?gat?atc?tac?aag?tac 491
Asp?Asn?Pro?Gln?Leu?Val?Ser?Glu?Tyr?Val?Asn?Asp?Ile?Tyr?Lys?Tyr
129 135 141
ctg?cga?gag?ctg?gag?gat?gcc?aac?aaa?gtc?aag?ccc?aga?tac?cta?gaa 539
Leu?Arg?Glu?Leu?Glu?Asp?Ala?Asn?Lys?Val?Lys?Pro?Arg?Tyr?Leu?Glu
145 151 157
ggc?caa?gta?att?aca?gga?aag?atg?aga?gca?att?ttg?att?gac?tgg?ctt 587
Gly?Gln?Val?Ile?Thr?Gly?Lys?Met?Arg?Ala?Ile?Leu?Ile?Asp?Trp?Leu
161 167 173
gtc?caa?gta?cac?ctc?cgc?ttc?act?ctg?ctt?caa?gaa?aca?ctg?tat?ctg 635
Val?Gln?Val?His?Leu?Arg?Phe?Thr?Leu?Leu?Gln?Glu?Thr?Leu?Tyr?Leu
177 183 189
act?gtt?gct?atc?att?gac?aga?ttt?ctc?cag?act?cag?agg?aat?ata?cca 683
Thr?Val?Ala?Ile?Ile?Asp?Arg?Phe?Leu?Gln?Thr?Gln?Arg?Asn?Ile?Pro
193 199 205
cgt?aac?aag?cta?cag?tta?gtt?ggt?gtg?act?gcc?atg?ttc?ata?gct?agt 731
Arg?Asn?Lys?Leu?Gln?Leu?Val?Gly?Val?Thr?Ala?Met?Phe?Ile?Ala?Ser
209 215 221
aaa?tat?gaa?gaa?atg?tat?tgc?cca?gaa?atc?ggg?gac?ttc?gca?tac?atc 779
Lys?Tyr?Glu?Glu?Met?Tyr?Cys?Pro?Glu?Ile?Gly?Asp?Phe?Ala?Tyr?Ile
225 231 237
aca?gac?aaa?gcc?tac?tca?aag?gca?gag?att?cgt?aaa?atg?gag?gtg?acc 827
Thr?Asp?Lys?Ala?Tyr?Ser?Lys?Ala?Glu?Ile?Arg?Lys?Met?Glu?Val?Thr
241 247 253
atg?ctg?aat?gag?ctg?ggc?ttc?aat?gta?tcc?tat?ccc?ctt?ccc?tta?cac 875
Met?Leu?Asn?Glu?Leu?Gly?Phe?Asn?Val?Ser?Tyr?Pro?Leu?Pro?Leu?His
257 263 269
ttc?ctg?cga?aga?aac?agc?aaa?gct?ggc?tct?gtt?gat?gct?tct?caa?cac 923
Phe?Leu?Arg?Arg?Asn?Ser?Lys?Ala?Gly?Ser?Val?Asp?Ala?Ser?Gln?His
273 279 285
acc?ttg?gca?aag?tac?ttg?atg?gaa?ctt?tgc?ttg?cca?gag?tac?agc?atg 971
Thr?Leu?Ala?Lys?Tyr?Leu?Met?Glu?Leu?Cys?Leu?Pro?Glu?Tyr?Ser?Met
289 295 301
tgc?cat?tac?aag?tca?tca?atg?att?gct?gca?tct?gct?ctc?tgc?ctt?tca 1019
Cys?His?Tyr?Lys?Ser?Ser?Met?Ile?Ala?Ala?Ser?Ala?Leu?Cys?Leu?Ser
305 311 317
ctt?aaa?ttg?ttg?gat?ggc?aat?aac?tgg?agt?gat?aca?ttg?act?ttc?tat 1067
Leu?Lys?Leu?Leu?Asp?Gly?Asn?Asn?Trp?Ser?Asp?Thr?Leu?Thr?Phe?Tyr
321 327 333
tct?cgc?tac?act?gaa?caa?cag?ctc?atg?cca?gtc?atg?tgc?aaa?atg?gca 1115
Ser?Arg?Tyr?Thr?Glu?Gln?Gln?Leu?Met?Pro?Val?Met?Cys?Lys?Met?Ala
337 343 349
tca?gtt?gta?gta?aag?agc?agt?agt?gcc?aag?caa?cag?gct?gta?aga?cag 1163
Ser?Val?Val?Val?Lys?Ser?Ser?Ser?Ala?Lys?Gln?Gln?Ala?Val?Arg?Gln
353 359 365
aag?tac?aaa?gcc?agc?aag?ttg?atg?aaa?att?agt?gag?att?ccc?cag?ctc 1211
Lys?Tyr?Lys?Ala?Ser?Lys?Leu?Met?Lys?Ile?Ser?Glu?Ile?Pro?Gln?Leu
369 375 381
aaa?tcg?aag?ctc?atc?aac?tcg?ctc?gca?gag?aag?agt?gcg?tct?tat?gca 1259
Lys?Ser?Lys?Leu?Ile?Asn?Ser?Leu?Ala?Glu?Lys?Ser?Ala?Ser?Tyr?Ala
385 391 397
tga 1262
*
ggtggtcgcc?atttaaaaag?taaatattgt?tcatgttgaa?agaatggggt?tgtttttgta 1322
catatttttt?taagacgcca?gttttttttt?ttcataagtt?ttcagattta?taaaatactg 1382
accaccagtt?tttttttttt?tttttaacaa?gagtagtttt?ggggggcagc?atgtaccccc 1442
cagagtgggt?ggtttcggca?tccctccatt?catcgtggtc?ttcattgtat?taaaataatt 1502
ttaggcagga?caaaaatggt?tatttgaaga?aaggaaccag?caaagaagta?tatgcaatgt 1562
cttgtaatgt?atatatgacc?tgtgataaac?ttttagttat?gttctgtatg?tatacctata 1622
taataaacca?gttttatcca?aaaaaaaaaa?aaaaaa 1658
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| CN 200610037559 CN1928085A (en) | 2006-09-07 | 2006-09-07 | Gene sequence of penaeus monodon cell periodic protein B |
| CNB2007100912290A CN100554421C (en) | 2006-09-07 | 2007-03-23 | The gene order of tigar prawn cell periodic protein B |
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| CN 200610037559 CN1928085A (en) | 2006-09-07 | 2006-09-07 | Gene sequence of penaeus monodon cell periodic protein B |
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