Molecular cloning, sequence analysis, and tissue expression of Smad3-like protein from Eriocheir sinensis
2018; Science Press; Volume: 25; Issue: 2 Linguagem: Inglês
10.3724/sp.j.1118.2018.17155
ISSN1005-8737
AutoresZhihuan Tian, Chuanzhen Jiao, Yongxu Cheng,
Tópico(s)Animal Genetics and Reproduction
ResumoPDF HTML阅读 XML下载 导出引用 引用提醒 中华绒螯蟹Smad3(EsSmad3)的cDNA克隆、序列分析及表达特征 DOI: 作者: 作者单位: 1. 广东韶关学院英东生命科学学院, 广东 韶关 512005;2. 上海海洋大学 农业部淡水水产种质资源重点实验室, 上海 201306 作者简介: 田志环(1971-),博士,副教授,研究方向:甲壳动物生长发育.E-mail:tianzhihuan@126.com 通讯作者: 中图分类号: S917 基金项目: 国家自然科学基金项目(31572635);科技部港澳台科技合作专项项目(2014DFT30270);上海市科学技术委员会科研项目(16DZ2281200);上海高校水产学高峰学科建设项目(2015-62-0908);上海市科技兴农推广项目[沪农科推字(2015)第1-7号];韶关学院生态学重点扶持学科(230079030101). Molecular cloning, sequence analysis, and tissue expression of Smad3-like protein from Eriocheir sinensis Author: Affiliation: 1. College of Yingdong Life Science, Shaoguan University, Shaoguan 512005, China;2. Key Laboratory of Freshwater Fishery Germplasm Resources, Ministry of Agriculture;Shanghai Ocean University, Shanghai 201306, China Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:为了研究Smad3(命名为)的cDNA全长序列2021 bp,包括36 bp的5'非翻译区(5'-UTR)、656 bp的3'非翻译区(3'-UTR)和编码442个氨基酸的开放阅读框。蛋白质结构域分析显示EsSmad3含有MH1和MH2两个特征性保守结构域。多序列比对显示,EsSmad3与人、斑马鱼、黑腹果蝇中的同源蛋白序列一致性分别为0.679、0.691、0.619。应用荧光定量RT-PCR技术分析在性成熟个体的肝胰腺、眼柄、表皮、卵巢、精巢、心脏、螯足、鳃、三角膜等组织中均有表达,其中眼柄和精巢中表达量较高,心脏和肝胰腺中表达量最低。在幼体不同蜕壳时期的不同部位的肌肉中,期和蜕壳后A~B期,但无显著的统计学差异(<0.05),蜕壳后A~B期开始表达量显著升高( mRNA水平在蜕皮间期C期显著高于蜕壳后A~B期,这种上调一直持续到蜕壳前晚前期D3-4。上述结果表明,参与了中华绒螯蟹蜕壳诱导的肌肉萎缩、生长及重建过程。 Abstract:In the present study, full length cDNA, encoding the ), was cloned using 3' Race and 5' Race techniques, and the sequence and structural analysis of was conducted using bioinformatics methods. The results showed that the full-length cDNA encoding consisted of 2021 bp nucleic acids in length, including a 5'-UTR of 36 bp, a 3'-UTR of 656 bp and an open reading frame (ORF) which encoded 442 amino acids. Analysis of the protein domain features showed that the deduced polypeptides contained two conservative domains characteristic of MH1 and MH2. Multiple sequence alignment revealed that the amino acid sequences of EsSmad3 have the 0.679, 0.691, and 0.619 identity with respectively. The tissue distribution of mRNA in sexually mature individuals and different muscle groups during the molt cycle in juvenile crabs, were analyzed using quantitative real-time PCR (qRT-PCR). In sexually mature crabs, the transcript was detected in the eyestalk, claw muscle, ovary, heart, hepatopancreas, epidermis, testicle, gill, and triangular membrane, and the expression level was relatively high in the eyestalk and testicle, and was low in the hepatopancreas and heart. In juvenile crabs, the transcript in different muscle groups was different depending on the molt stage. In walking leg muscles, the expression level was higher in inter-molt C stage than in the later pre-molt D3-4 and post-molt A-B stages, but there was no statistically significant difference ( expression level decreased rapidly in the pre-molt D3-4 stage (<0.05) and increased in the post-molt A-B stage, lasting to the inter-molt C stage ( expression level was much higher in the inter-molt C stage than in the post-molt A-B stage (stage. These results suggest that the expression of transcript in different muscle groups was related to the molt stage of is involved in muscle atrophy, growth, and rebuilding during the molt cycle of 参考文献 相似文献 引证文献
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