缢蛏生长因子受体结合蛋白2基因克隆、时空表达及SNP筛查

赵家熙; 崔宝月; 董迎辉; 姚韩韩; 林志华

doi:10.3969/j.issn.0253-4193.2018.02.009

缢蛏生长因子受体结合蛋白2基因克隆、时空表达及SNP筛查

doi: 10.3969/j.issn.0253-4193.2018.02.009

1.
浙江万里学院浙江省水产种质资源高效利用技术研究重点实验室, 浙江宁波 315100
2.
上海海洋大学水产与生命学院, 上海 201306

基金项目: 国家现代贝类产业技术体系项目（CARS-49）；浙江省农业新品种选育重大科技专项（2016C02055-9）；国家水产种质资源平台运行服务项目（2018DKA30470）；宁波市自然科学基金项目（2016C70002）项目资助。

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出版历程
- 收稿日期: 2017-07-11

Cloning, spatiotemporal expression and SNPs identification of GRB2 gene in Sinonovacula constricta

1.
Key Laboratory of Aquatic Germplasm Resource of Zhejiang Province, Zhejiang Wanli University, Ningbo 315100, China
2.
College of Fishery and Life Sciences, Shanghai Ocean University, Shanghai 201306, China

摘要

摘要: 为了探索生长因子受体结合蛋白2（GRB2）在缢蛏生长发育中的作用，本实验基于缢蛏转录组文库，利用SMART RACE技术克隆缢蛏GRB2（Sc-GRB2）基因的cDNA全长序列，分析其在不同组织和发育时期的表达差异，并在外显子中进行了SNP位点筛选。结果表明，Sc-GRB2基因cDNA全长1 223 bp，开放阅读框711 bp，编码236个氨基酸；氨基酸序列比对发现，缢蛏与文蛤、泥蚶等双壳贝类同源性较高（64%、59%），而与其他物种的同源性为45%~58%，表明GRB2基因比较保守；不同组织的荧光定量PCR （qRT-PCR）结果显示，Sc-GRB2基因在缢蛏7个组织中均有表达，其中足中的表达量极显著地高于其他6个组织（P<0.01）；在8个发育时期的表达差异分析发现，该基因在稚贝期表达量极显著地高于其他7个时期（P<0.01）。SNP位点筛选结果表明，在Sc-GRB2基因的外显子区域共发现11个SNP位点。
- 缢蛏 /
- 生长因子受体结合蛋白2 /
- 克隆 /
- SNP
Abstract: To explore the molecular structure and biological function of GRB2 in Sinonovacula constricta(Sc-GRB2), its cDNA was cloned by SMART RACE techniques, then the bioinformatics, SNPs in exon and expression profiles in different tissues and developmental stages were analyzed. The results indicated that the full length cDNA of Sc-GRB2 gene was 1 223 bp, containing a complete 711 bp ORF encoding 236 amino acids. Comparisons of animal acid sequence, Sc-GRB2 has high homology with bivalves of Meretrix meretrix and Tegillarca granosa (64% and 59%). Sc-GRB2 has homologous with others share 45%-58% similarity, it indicate GRB2 is relatively conservative. The result of qRT-PCR in different tissues showed that Sc-GRB2 expressed in all seven tissues, the expression of foot was extremely significantly higher than others (P<0.01). The relative expression in different stages revealed that the expression of juvenile clams was extremely significantly higher than other stages (P<0.01). A total of 11 SNPs in the exon of Sc-GRB2 were identified.
- Sinonovacula constricta /
- GRB2 /
- gene cloning /
- SNP

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