CN102021172A - Tilapia mossambica white blood cell inducible cAMP early repressor (ICER) gene sequence and applications thereof - Google Patents
Tilapia mossambica white blood cell inducible cAMP early repressor (ICER) gene sequence and applications thereof Download PDFInfo
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Abstract
The invention discloses a tilapia mossambica white blood cell inducible cAMP early repressor (ICER) gene sequence and applications thereof. The overall length of the gene cDNA sequence is 569bp, and the gene cDNA sequence comprises one complete open reading frame, and codes 111 amino acid; the sequence shows the basic structural feature of ICER II gamma, i.e. the sequence only has cAMP response element regulation and control factor (CREM) leucine zipper DNA combination structural area DBDII, and has no exon gamma. The gene plays an important role in multiple cell physiological activities, and is applied in tilapia mossambica disease-resistant breeding.
Description
Technical field
The present invention relates to tilapia white corpuscle cAMP and induce early stage supressor (inducible cAMP early repressor, ICER) gene order, more specifically relate to tilapia white corpuscle SSH cDNA library order-checking before and after the immunity, analyze the tilapia ICER full length cDNA sequence, aminoacid sequence, the spatial and temporal expression situation that obtain, the invention still further relates to its purposes.
Background technology
The cAMP signal transduction path has important biological function in a plurality of fields such as reproductive system, neural system, immunity system and endocrine system of higher animal.ICER is a kind of transcription inhibition factor of cAMP response element regulatory factor genes encoding, can suppress the expression of self by the automatically negative regulation mechanism of itself, and the human ICER that has reported at present comprises ICER I, ICERI γ, ICER II, and ICER II γ.ICER participates in numerous cell physiological activities, be the daily cycle variation as pineal gland ICER, suppress the growth of prostate cancer cell, N-methyl D-aspartic acid receptor antagonists etc. die, promote aspects such as corpus luteumization, corticotrope cycle and regulation of secretion to have vital role at inhibition growth of tumour cell, neurocyte generation and accent.At present, the research of relevant fish ICER report still belongs to blank, and tilapia ICER gene order is not appeared in the newspapers yet.
Summary of the invention
The object of the present invention is to provide tilapia peripheral blood leucocyte cAMP to induce the gene order of early stage supressor, aminoacid sequence and spatial and temporal expression situation, analytical procedure is easy.
The invention still further relates to tilapia peripheral blood leucocyte cAMP and induce the application of early stage supressor gene order in the disease-resistant seed selection of tilapia.
For realizing above-mentioned task, the present invention has adopted following technical measures:
Utilize the inhibition difference to subtract hybridization technique (SSH), with tilapia white corpuscle cDNA before and after the streptococcicosis vaccine immunity is material, tilapia immunity front and back forward and reverse two cDNA subtracted libraries have successfully been made up, by the library sequencing result being unloaded analyses such as body, sequence assembly, homology retrieval, obtain the ICER full length cDNA sequence.This sequence length overall 569bp comprises 1 complete open reading frame, the open reading frame of the long 336bp of this gene cDNA, 111 amino acid of encoding.5 ' end non-coding region 119bp, 3 ' end non-coding region 114bp, the long 30bp of PolyA tail.The basic structural feature of ICER II γ is revealed in sequence table, cAMP response element regulatory factor (CREM) leucine zipper DNA binding domains DBD II is promptly only arranged, and do not have exon γ.Motif (motif) search shows that ICER II γ/T contains N-glycosylation site NKTL (92); protein kinase C phosphorylation site TRK (53); 2 tyrosine protein kinase II phosphorylation site TGDE; TLIE (4; 94), N-acylations site GLAQSI (27), leucine zipper transcription factor structural domain sequence label RLMKNREAARECRRK (59); leucine zipper motif sequence LENRVAVLENQNKTLIEELKAL (81), nuclear localization sequence RKREVRLMKNREAARECRRKKKE (53-70).Only obtain 1 homologous sequence through the BLASTN retrieval, i.e. the cAMP response element regulatory factor mRNA of Atlantic salmon supposition.Adopt the real time fluorescent quantitative method to immunity with attack poison back Buddhist nun difficult to understand and hybridize tilapia, bolti, Oreochromis aureus and gift tilapia ICER and carry out the spatial and temporal expression analysis.
Gene ICER cDNA sequence:
GAGGAAAGGAGAAGCGGGAGCAAAGGAGATACAAGTAAAGCAGAAAAGGATAGTGGTAGAGAAGAGTTTTTGTAGTGAGG
TGTCTCTCTACCGTGGTGCTGGATTATTGGAAACCAGAGATGGCTGTAACTGGGGATGAGACTGAGTCAGCCGCTACTGG
AGACATCCCCGCCTACCAGCTTCGTTCACCAAACTCGGGCTTGGCTCAGAGCATTGTAATGGCTGCGTCTCCAGGCAGCA
TGCAGAGCCCCTCATCACAGCACGCCGAGGAGATCACCCGCAAGAGGGAGGTCCGGCTGATGAAAAACAGGGAAGCAGCT
CGCGAGTGCCGCAGGAAAAAGAAAGAGTATGTCAAATGTCTGGAAAACCGCGTGGCTGTGTTGGAAAACCAAAACAAGAC
CTTGATTGAAGAGCTAAAAGCACTAAAGGACATTTACTGCCACAAAGCTGAGTAGCAGTGGCTGCTTGTGGTCATGGGCT
GAAGACAGTTTACAAACTTCTACAGCAAAATAAGACAAAACAAAACAAAACAAAAAACACAAAAAAAAAAAAAAAAAAAA
AAAAAAAAA
The aminoacid sequence of gene ICER proteins encoded:
MAVTGDETESAATGDIPAYQLRSPNSGLAQSIVMAASPGSMQSPSSQHAEEITRKREVRLMKNREAARECRRKKKEYVKC
LENRVAVLENQNKTLIEELKALKDIYCHKAE
Tilapia white corpuscle cAMP induces the application of early stage supressor gene order in the disease-resistant seed selection of tilapia.
By ICER genetic expression real-time analysis, attack all in various degree downward modulations of ICER expression amount of brain, liver, kidney and the four kinds of tissues of spleen of the back four kinds of fishes of poison, it all is first downward modulations that brain, liver, kidney and the spleen of the back four kinds of fishes of immunity are organized the ICER expression amount for four kinds, transfers rise behind the 48h again to.In addition, attack whole expression amount of poison back ICER and immunity later stage ICER and go up modulation factor of amplitude modulation and hybridize to be in tilapia, bolti, Oreochromis aureus and the gift tilapia successively Buddhist nun difficult to understand and descend, with testing laboratory's streptococcus infection abilities of four kinds of tilapias with produce and go up disease resistance and be proportionate.
Beneficial effect of the present invention:
The ICER gene is to find first tilapia, at present humanly tilapia the understanding and the understanding of this gene is only limited to the research-on-research research that the inventor does.ICER has consequence in tilapia body life adjusting activity, by analyzing its spatial and temporal expression situation, explore the relation of the disease-resistant difference of ICER and different varieties tilapia, for the disease-resistant seed selection of tilapia provides molecule marker to instruct the disease-resistant seed selection of tilapia.
Description of drawings
Total RNA denaturing formaldehyde electrophoresis result before and after the immunity of Fig. 1 tilapia.
Fig. 2 bolti β-actin gene copy logarithmic amplification typical curve.
Fig. 3 bolti ICER gene copy logarithmic amplification typical curve.
The back four kinds of tilapia cerebral tissue ICER expression levels of Fig. 4 streptococcus iniae vaccine immunity.
The back four kinds of tilapia hepatic tissue ICER expression levels of Fig. 5 streptococcus iniae vaccine immunity.
The back four kinds of tilapia spleens of Fig. 6 streptococcus iniae vaccine immunity are organized the ICER expression level.
The back four kinds of tilapia nephridial tissue ICER expression levels of Fig. 7 streptococcus iniae vaccine immunity.
Fig. 8 Streptococcus iniae infects back four kinds of tilapia cerebral tissue ICER expression levels.
Fig. 9 Streptococcus iniae infects back four kinds of tilapia hepatic tissue ICER expression levels.
Figure 10 Streptococcus iniae infects back four kinds of tilapia spleens and organizes the ICER expression level.
Figure 11 Streptococcus iniae infects back four kinds of tilapia nephridial tissue ICER expression levels.
Indicate (Fig. 4～Figure 11): sample encoded the 1st bit representation tissue (L: liver, K: pronephridiostome, S: spleen, B: brain); The kind of the 2nd bit representation fish (1: Ji Fu, 2: Ao Liya, 3: Buddhist nun sieve, 4: Buddhist nun's hybridization difficult to understand); The 3rd bit representation time point (1:0h, 2:6h, 3:12h, 4:24h, 5:48h).
Embodiment
1, the structure of the extraction of RNA and SSH subtracted library
Tilapia is divided into two groups and raises in the small-sized net cage of fish pond.Immune group tilapia suis vaccine peritoneal immunity (10
9Cell/ml, the 0.5ml/ tail), two all pneumoretroperitoneum immunity (10
9Cell/ml, the 0.5ml/ tail) booster immunization is once.Booster immunization is two groups of tilapia tail vein bloods (15IU anticoagulant heparin 1ml blood) 14ml after one day, slowly be added to isopyknic lymphocyte separation medium surface again after adding the dilution of PBS twice, 18 ℃, 500g * 50min, leukocytic cream in the middle of careful the absorption, the PBS of adding and blood sampling volume equivalent, the centrifugal 20min of 500g collects white corpuscle, repeat twice, it is resuspended to add an amount of PBS.Wright's staining is identified and the cell counting count board counting.Per 10
7Individual white corpuscle adds 1ml Trizol reagent, and concuss is to not having visible cell group, room temperature effect 5min; Room temperature effect 5min behind every milliliter of Trizol adding 0.2ml chloroform concuss 30s, 4 ℃, 12000g * 15min; The careful upper strata water of drawing, every milliliter of Trizol adds 0.5ml Virahol-20 ℃ precipitation 30min, 4 ℃, 12000g * 10min; Abandon supernatant, every milliliter of Trizol adds 4 ℃ of 1ml 75% ethanol, twice of 7500g * 5min washing precipitation; Drying at room temperature 15min adds an amount of DEPC water dissolution (55 ℃ of water-bath hydrotropy 5min).The nucleic acid-protein determinator detects OD260/OD280, and tilapia A260/A280 is 1.97 before the immunity, and total amount is 123ug; Immunity back tilapia A260/A280 is 2.0, and total amount is 150ug.Through the sex change electrophoresis detection, 28S and 18S RNA banding pattern are high-visible, and ratio is (Fig. 1) suitably, proves that total RNA quality is good, purity is high.The mRNA that obtains is carried out freeze-drying concentrate raising concentration.
According to the PCR-Select of U.S. CLONTECH company
TMCDNA Subtraction Kit explanation makes up the SSH subtracted library, and concrete operations are as follows: with tilapia white corpuscle cDNA after the immunity is the Tester group, and normal tilapia white corpuscle cDNA is the Driver group.Be divided into two parts after Tester cDNA enzyme is cut, finish Tester-1 and Tester-2cDNA after connecting two kinds of different joint Adptor1 and Adaptor2R respectively.Enzyme cut the Driver cDNA of back preparation and the Tester cDNA after joint is connected carries out two-wheeled and hybridizes.The first round, excessive Driver cDNA is added to every part of Tester cDNA, behind 98 ℃ of sex change 1.5min, and 68 ℃ of hybridization 8h.Difference expression gene cDNA fragment does not form strand with Driver cDNA hybridization.And then carry out second and take turns hybridization, without sex change rapidly with two parts of sample mix of hybridization for the first time, add the Driver cDNA after the sex change simultaneously, 68 ℃ are continued hybridization 10h behind the mixing, and the strand cDNA of differential expression anneals again and is formed with the double-stranded cDNA of different connector end.The hybridization product is again through two-wheeled PCR reaction.First round PCR adopts the consensus sequence primer PCR Primer 1 of Adaptor1 and Adaptor2R.The amplification parameter is: 94 ℃, and 25s; 94 ℃, 10s; 66 ℃, 30s; 72 ℃, 1.5min; 25 circulations.The cDNA fragment of having only hybridization formation two ends to have different joints can increase.Second that take turns that PCR adopts is sleeve type PCR primer (Nested primer1 and Nested Primer2R), and the amplification parameter is: 94 ℃, and 10s; 68 ℃, 30s, 72 ℃, 1.5min, 15 circulations.With normal tilapia is that Tester group, immunity back tilapia are the Driver group, carries out the inhibition difference by above-mentioned forward crossover operation step and subtracts hybridization, to obtain difference expression gene.Positive and negative difference subtract in the PCR product (50ul altogether) get 1ul and pGEM-T carry out T the A cloning vector spend the night for 4 ℃ and be connected.Connect among the product transformed into escherichia coli JM109.Get 20ul and be coated on the LB/Amp/IPTG/X-Cal flat board and carry out the screening of blue hickie, picking white is cloned in the LB substratum that contains Amp (50ug/ml) 37 ℃, 150rpm shaking culture 16h.Original library glycerol adding to 20% ,-80 ℃ of preservations.
2, sequential analysis
Positive and negative library random choose positive colony checks order, carry out nucleic acid sequence homology retrieval and comparison by blast program at the nr of international gene database GenBank+EMBL+DDBJ+PDB database and est database, 56 effective est sequences of acquisition and animal ICER cDNA sequence similarity are very high from positive storehouse.Utilize DNASTAR/SeqMan software that 56 EST are spliced, the contig that obtains is at last respectively searched for nr storehouse and swissport storehouse with BLASTN and BLASTP as target sequence on NCBI, seek homology sequence, ORF analyzes, and determines that finally gained contig is the ICER full length cDNA sequence.This sequence length overall 569bp comprises 1 complete open reading frame, 111 amino acid of encoding.The basic structural feature of ICER II γ is revealed in sequence table, cAMP response element regulatory factor (CREM) leucine zipper DNA binding domains DBD II is promptly only arranged, and do not have exon γ.Motif (motif) search shows that ICER II γ/T contains N-glycosylation site NKTL (92); protein kinase C phosphorylation site TRK (53); 2 tyrosine protein kinase II phosphorylation site TGDE; TLIE (4; 94); N-acylations site GLAQSI (27); leucine zipper transcription factor structural domain sequence label RLMKNREAARECRRK (59); leucine zipper motif sequence LENRVAVLENQNKTLIEELKAL (81); nuclear localization sequence RKREVRLMKNREAARECRRKKKE (53-70), the calling sequence called after ICERII γ/T of institute (the GenBank number of landing is EU593895).
3, the ICER spatial and temporal expression is analyzed
A, primer: ICER upstream primer: 5 '-AGTCAGCCGCTACTGGAGACA-3 ' and downstream primer: 5 '-CTGGAGACGCAGCCATTACA-3 ', amplified fragments 90bp.β-Actin upstream primer: 5 '-AACAACCACACACCACACATTTC-3 ' and downstream primer: 5 '-TGTCTCCTTCATCGTTCCAGTTT-3 ' amplified fragments 134bp.
B, Buddhist nun difficult to understand hybridize that tilapia, bolti, Oreochromis aureus and gift tilapia are divided into immune group and challenge test group (50 tail/group) is raised respectively in the pond small-sized net cage that separates.Immune group tilapia abdominal injection suis vaccine carries out immunity (1.0 * 10
9Cell/ml, 0.5ml/ tail), get 3 tails test fish at random respectively at (0d) before the immunity and immunity back 1d, 3d, 5d, 7d, dissection is got its brain, liver, spleen and pronephridiostome and is organized in the liquid nitrogen and preserves.Challenge test group tilapia abdominal injection suis carries out artificial challenge (1.0 * 10
8Cell/ml, the 0.5ml/ tail), each is organized respectively at (0h) before infecting and infection back 6h, 12h, 24h, 48h and gets 3 tail survival test fishes at random, and dissection is got its brain, liver, spleen and nephridial tissue and is preserved in liquid nitrogen.
C, RNA extracting and reverse transcription are pressed mark and take out preservation tissue and grind into powder from liquid nitrogen, extract total RNA and select for use the reverse transcription universal primer to carry out the synthetic cDNA of reverse transcription.
D, amplification efficiency detect, with cDNA concentration gradient Lg value to C
tThe value mapping, the interior mark β-actin typical curve (Fig. 2) of foundation and the typical curve (Fig. 3) of ICER gene, its relation conefficient is respectively 0.999 and 1.000, and dependency is fine.According to formula LgXo=-Lg (1+E
x) * C
t+ LgN, ICER quantitative PCR reaction efficiency is 0.99947.
E, ICER detection by quantitative adopt two-step approach to carry out the real-time fluorescence quantitative PCR reaction, and reaction conditions is the first step, 95 degree 10s, second step, 95 degree, 5s; 60 degree, 34s, 40 circulations.Reaction is pressed Δ Δ CT value analytical procedure after finishing, and uses 7500 Software v2.0.1 (ABI) analytical data, making spatial and temporal expression collection of illustrative plates (Fig. 4～Figure 11), sample encoded the 1st bit representation tissue (L: liver, K: pronephridiostome, S: spleen, B: brain), the kind of the 2nd bit representation fish (1: Ji Fu, 2: Ao Liya, 3: Buddhist nun sieve, 4: Buddhist nun's hybridization difficult to understand), the 3rd bit representation time point (1:0d, 2:1d, 3:3d, 4:5d, 5:7d); Fig. 5 sample encoded the 1st bit representation tissue (L: liver, K: pronephridiostome, S: spleen, B: brain), the kind of the 2nd bit representation fish (1: Ji Fu, 2: Ao Liya, 3: Buddhist nun sieve, 4: Buddhist nun's hybridization difficult to understand), the 3rd bit representation time point (1:0h, 2:6h, 3:12h, 4:24h, 5:48h).By ICER genetic expression real-time analysis, attacking malicious and immune back tilapia liver, brain, spleen and preceding nephridial tissue ICER gene expression amount all descends in various degree, but rising significantly appears again in four kinds of tilapias ICER expression amount behind 7d that immunity is handled, near handling preceding expression level and the trend that surpasses being arranged.Four kinds of tilapia ICER expression ratios are found, under the normal circumstances Buddhist nun difficult to understand hybridize tilapia ICER liver, brain, spleen and before in the nephridial tissue expression amount all be significantly higher than other 3 kinds of tilapias, attack whole expression amount that poison back Buddhist nun difficult to understand hybridizes tilapia ICER also apparently higher than other 3 kinds of tilapias, immunity later stage Buddhist nun difficult to understand is hybridized tilapia ICER and is gone up that modulation factor of amplitude modulation is also bright to be higher than other 3 kinds of tilapias.By computational analysis, attack whole expression amount of poison back ICER and immunity later stage ICER and go up modulation factor of amplitude modulation and hybridize tilapia, bolti, Oreochromis aureus and gift tilapia Buddhist nun difficult to understand and be successively and descend.Suis is attacked poison and found that Buddhist nun difficult to understand hybridizes tilapia, bolti, Oreochromis aureus and gift tilapia mortality ratio and be successively and rise, illustrate that four kinds of tilapia streptococcus infection abilities are decline successively, this result is consistent with the comprehensive resistance against diseases of producing these four kinds of tilapias performances in the breed in recent years.Attack poison or immunity back change of Expression trend, four kinds of tilapia ICER differential expressions and anti-infection ability comparative result in conjunction with ICER, find that ICER gene expression amount and tilapia disease resistance have inherent positive correlation, promptly the ICER expression amount is high more within the specific limits, helps the infection of pathogenic bacterias such as tilapia body opposing suis more.
F, four kinds of tilapias infect the suis comparison test: Buddhist nun difficult to understand is hybridized tilapia, bolti, Oreochromis aureus and gift tilapia and is divided into streptococcal infection test group and blank group (body weight 40-60g, 50 tail/groups), infective dose is a bolti medium lethal dose 2.0 * 10
7CFU, control group abdominal injection equal-volume physiological saline.Tilapia was raised and train for 1 week in the large volume water vat earlier in test.Infection experiment is carried out in the 45L water tank, every 2d changes water 2/3, and water temperature is between 22-29 ℃, and feed intake at twice by body weight 2% every day, and incidence observed statistics, utilize the chicken blood meida that tilapia brain, liver, spleen and the kidney of being at death's door carried out the bacterium isolation identification simultaneously.Observations statistics by 25 days, four kinds of tilapias all in various degree appearance death, mainly wherein at 4-6 days, death stops after 15 days.Through the bacterium isolation identification, all be at death's door tilapia brain, liver, spleen and kidneys all are separated to infection strain CMS005, and the death condition of four kinds of tilapias sees Table 1.
Necrology behind four kinds of tilapias infection of table 1 suis
Therefore, can realize the disease-resistant seed selection of tilapia by the ICER differential expression between more different tilapia kinds or the same breed Different Individual.
Sequence table
CAMP induces early stage supressor (ICER) sequence table
＜110〉Guangxi Zhuang Autonomous Region Aquatic Institute
＜120〉tilapia white corpuscle cAMP induces early stage supressor gene order and purposes
<160>1
<210>1
<211>569
<212>DNA
＜213〉tilapia kind (Oreochromis sp.)
<220>
<221>CDS
<222>(120)...(455)
<400>1
gaggaaagga?gaagcgggag?caaaggagat?acaagtaaag?cagaaaagga?tagtggtaga?60
gaagagtttt?tgtagtgagg?tgtctctcta?ccgtggtgct?ggattattgg?aaaccagag 119
atg?gct?gta?act?ggg?gat?gag?act?gag?tca?gcc?gct?act?gga?gac?164
M A V T G D E T E S A A T G D
5 10 15
atc?ccc?gcc?tac?cag?ctt?cgt?tca?cca?aac?tcg?ggc?ttg?gct?cag?209
I P A Y Q L R S P N S G L A Q
20 25 30
agc?att?gta?atg?gct?gcg?tct?cca?ggc?agc?atg?cag?agc?ccc?tca?254
S I V M A A S P G S M Q S P S
35 40 45
tca?cag?cac?gcc?gag?gag?atc?acc?cgc?aag?agg?gag?gtc?cgg?ctg?299
S Q H A E E I T R K R E V R L
50 55 60
atg?aaa?aac?agg?gaa?gca?gct?cgc?gag?tgc?cgc?agg?aaa?aag?aaa?344
M K N R E A A R E C R R K K K
65 70 75
gag?tat?gtc?aaa?tgt?ctg?gaa?aac?cgc?gtg?gct?gtg?ttg?gaa?aac?389
E Y V K C L E N R V A V L E N
80 85 90
caa?aac?aag?acc?ttg?att?gaa?gag?cta?aaa?gca?cta?aag?gac?att?434
Q N K T L I E E L K A L K D I
95 100 105
tac?tgc?cac?aaa?gct?gag?452
Y C H K A E
110
tagcagtggc?tgcttgtggt?catgggctga?agacagttta?caaacttcta?cagcaaaata?512
agacaaaaca?aaacaaaaca?aaaaacacaa?aaaaaaaaaa?aaaaaaaaaa?aaaaaaa 569
<210>2
<211>111
<212>PRT
＜213〉tilapia kind (Oreochromis sp.)
<400>2
M A V T G D E T E S A A T G D
5 10 15
I P A Y Q L R S P N S G L A Q
20 25 30
S I V M A A S P G S M Q S P S
35 40 45
S Q H A E E I T R K R E V R L
50 55 60
M K N R E A A R E C R R K K K
65 70 75
E Y V K C L E N R V A V L E N
80 85 90
Q N K T L I E E L K A L K D I
95 100 105
Y C H K A E
110
Claims (2)
1. tilapia white corpuscle cAMP induces early stage supressor gene order, it is characterized in that: the aminoacid sequence of gene ICER cDNA sequence and gene ICER proteins encoded; The open reading frame of the long 336bp of this gene cDNA, 111 amino acid of encoding, 5 ' end non-coding region 119bp, 3 ' end non-coding region 114bp, the long 30bp of PolyA tail;
Gene ICER cDNA sequence:
GAGGAAAGGAGAAGCGGGAGCAAAGGAGATACAAGTAAAGCAGAAAAGGATAGTGGT
AGAGAAGAGTTTTTGTAGTGAGGTGTCTCTCTACCGTGGTGCTGGATTATTGGAAACCA
GAGATGGCTGTAACTGGGGATGAGACTGAGTCAGCCGCTACTGGAGACATCCCCGCCTA
CCAGCTTCGTTCACCAAACTCGGGCTTGGCTCAGAGCATTGTAATGGCTGCGTCTCCAG
GCAGCATGCAGAGCCCCTCATCACAGCACGCCGAGGAGATCACCCGCAAGAGGGAGGT
CCGGCTGATGAAAAACAGGGAAGCAGCTCGCGAGTGCCGCAGGAAAAAGAAAGAGTA
TGTCAAATGTCTGGAAAACCGCGTGGCTGTGTTGGAAAACCAAAACAAGACCTTGATT
GAAGAGCTAAAAGCACTAAAGGACATTTACTGCCACAAAGCTGAGTAGCAGTGGCTGC
TTGTGGTCATGGGCTGAAGACAGTTTACAAACTTCTACAGCAAAATAAGACAAAACAAA
ACAAAACAAAAAACACAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
The aminoacid sequence of gene ICER proteins encoded:
MAVTGDETESAATGDIPAYQLRSPNSGLAQSIVMAASPGSMQSPSSQHAEEITRKREVRLMKNREAARECRRKKKEYVKC
LENRVAVLENQNKTLIEELKALKDIYCHKAE。
2. tilapia white corpuscle cAMP induces the application of early stage supressor gene order in the disease-resistant seed selection of tilapia.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103874502A (en) * | 2011-09-30 | 2014-06-18 | 遗传工程与生物技术中心 | Amino acid sequences for controlling pathogens |
| CN109504683A (en) * | 2018-11-13 | 2019-03-22 | 广西壮族自治区水产科学研究院 | The galectin-3 gene order and its cloning process of Tilapia mossambica |
| CN111690613A (en) * | 2020-07-07 | 2020-09-22 | 中国科学院水生生物研究所 | Tilapia gilvata brain nerve cell line, transfection method, culture method and application thereof |
-
2010
- 2010-06-23 CN CN 201010207325 patent/CN102021172A/en active Pending
Non-Patent Citations (3)
| Title |
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| 《Mol Biol Rep》 20090901 ming chen et al. Sequence and expression analysis of the gene encoding inducible cAMP early repressor in tilapia 第37卷, 第5期 2 * |
| 《中国预防兽医学报》 20020131 柴家前等 罗非鱼链球菌的分离鉴定 第24卷, 第1期 2 * |
| 《大连水产学院学报》 20091031 李超等 用链球菌疫苗免疫罗非鱼前后其差异表达基因的鉴定与分析 第24卷, 第5期 2 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103874502A (en) * | 2011-09-30 | 2014-06-18 | 遗传工程与生物技术中心 | Amino acid sequences for controlling pathogens |
| CN103874502B (en) * | 2011-09-30 | 2015-11-25 | 遗传工程与生物技术中心 | For controlling the aminoacid sequence of pathogen |
| CN109504683A (en) * | 2018-11-13 | 2019-03-22 | 广西壮族自治区水产科学研究院 | The galectin-3 gene order and its cloning process of Tilapia mossambica |
| CN111690613A (en) * | 2020-07-07 | 2020-09-22 | 中国科学院水生生物研究所 | Tilapia gilvata brain nerve cell line, transfection method, culture method and application thereof |
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