凡纳滨对虾核自身抗原精子蛋白基因克隆及其表达
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上海海洋大学 水产与生命学院 上海 201306

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颜婕(1991-),女,硕士研究生,主要从事海洋生物发育生物学和分子生物学研究.E-mail:yanjie106@163.com

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S917

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上海市教育委员会重点科研创新项目(12ZZ160).


Molecular cloning and expression analysis of the gene encoding nuclear autoantigenic sperm protein from white prawn (Litopenaeus vannamei)
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College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China

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    摘要:

    利用cDNA末端快速扩增技术克隆出凡纳滨对虾(基因cDNA全长2258 bp,其中包括92 bp 5’端非编码区,147 bp 3’端非编码区及2019 bp开放阅读区,编码673个氨基酸,预测分子量为74.18 kD。该序列已提交至GenBank,序列号为KT274811。在NCBI上进行序列比对后发现,其与斑节对虾( mRNA相对表达水平,结果表明,在卵巢中的表达量高于肝胰腺中的表达量,且在Ⅱ期表达量最高,Ⅲ期表达量最低。此外,通过构建系统进化树比较了凡纳滨对虾NASP与其他物种间的遗传距离。实验结果为进一步研究该基因在凡纳滨对虾卵巢发育中的作用提供了依据。

    Abstract:

    Many genes that play roles in shrimp vitellogenesis, such as that encoding nuclear autoantigenic sperm protein (NASP), have been identified by suppression subtractive hybridization. In shrimp, NASP plays an important role in ovarian maturation. In this study, the using the rapid amplification of cDNA ends technique. The full-length cDNA of consisted of 2258 bp with a 92-bp 5'-untranslated region (UTR), a 174-bp 3'-UTR, and a 2019-bp open reading frame, which encoded a protein of 673 amino acids with a predicted isoelectric point of 4.46 and a predicted molecular weight of 74.18 kDa. The sequence has been submitted to GenBank under the accession number KT274811. The putative protein had two conserved regions:SHNi-TPR and TPR_2. A sequence alignment analysis revealed that the Penaeus monodon (GenBank accession number:FJ040859.1). We constructed a phylogenetic tree based on NASP sequences to evaluate the evolutionary relationships of NASPs between L. vannamei was in the same branch as that of were investigated in the ovary and hepatopancreas of at different overy developmental stages by real-time fluorescent quantitative PCR. The results showed that transcripts were present at all stages, with the highest levels at the second stage and the lowest levels at the third stage. The NASP transcript levels were higher in the ovary than in the hepatopancreas, in which the transcript level was negligible. These results provide the basis for further research on the role of NASP in ovarian development in .

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颜婕,刘红,方昱,蔡生力.凡纳滨对虾核自身抗原精子蛋白基因克隆及其表达[J].中国水产科学,2016,23(5):1073-1079
YAN Jie, LIU Hong, FANG Yu, CAI Shengli. Molecular cloning and expression analysis of the gene encoding nuclear autoantigenic sperm protein from white prawn (Litopenaeus vannamei)[J]. Journal of Fishery Sciences of China,2016,23(5):1073-1079

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  • 在线发布日期: 2016-09-05
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