中国明对虾BMP-7基因的克隆及表达模式分析  

Cloning and Expression of FcBMP-7 Gene in Chinese Shrimp Fenneropenaeus chinensis

作　　者：高娜 卢霞 闫允君 孔杰[2,3] 孟宪红 李旭鹏[2,3] 隋娟 陈宝龙[2,3] 曹宝祥 高焕[1] 栾生 GAO Na;LU Xia;YAN Yunjun;KONG Jie;MENG Xianhong;LI Xupeng;SUI Juan;CHEN Baolong;CAO Baoxiang;GAO Huan;LUAN Sheng(Jiangsu Key Laboratory of Marine Biotechnology,School of Marine Science and Fisheries,Jiangsu Ocean University,Lianyungang 222005,China;Key Laboratory for Sustainable Development of Marine Fisheries,Ministry of Agriculture and Rural Affairs,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China;Laboratory for Marine Fisheries Science and Food Production Processes,Pilot National Laboratory for Marine Science and Technology(Qingdao),Qingdao 266071,China;Research and Development Center for Efficient Utilization of Coastal Biological Resources,Yantai Institute of Coastal Zone Research,Chinese Academy of Sciences,Yantai 264003,China)

机构地区：[1]江苏海洋大学海洋科学与水产学院,江苏省海洋生物技术重点试验室,江苏连云港222005 [2]中国水产科学研究院黄海水产研究所,农业农村部海洋渔业可持续发展重点实验室,山东青岛266071 [3]青岛海洋科学与技术国家实验室,海洋渔业科学与食物产出过程功能实验室,山东青岛266071 [4]中国科学院烟台海岸带研究所,沿海生物资源高效利用研究开发中心,山东烟台264003

出　　处：《水产科学》2023年第3期347-356,共10页Fisheries Science

基　　金：国家重点研发计划资助项目(2018YFD0901302);国家自然科学基金联合基金资助项目(U1706203);山东省农业良种工程项目(2019LZGC014).

摘　　要：为探明转化生长因子-β (TGF-β)超蛋白家族的成员之一骨形态发生蛋白7 (BMP-7)基因在中国明对虾肌肉发生和生长过程中的表达模式,通过cDNA末端快速扩增技术得到了中国明对虾BMP-7基因(FcBMP-7基因)2003 bp的全长cDNA,包含466 bp的5′非翻译区和295 bp的3′非翻译区,1242 bp的开放阅读框共编码413个氨基酸。利用实时荧光定量PCR技术分析FcBMP-7基因在中国明对虾幼体原肠期、无节幼体期、溞状幼体期、糠虾幼体期、仔虾期5个不同发育时期肌肉组织中的表达规律,结果显示,该基因在幼体发育的各时期均有表达,原肠期表达量最低,从无节幼体期开始表达量显著增加(P<0.05),糠虾期表达水平最高,表明FcBMP-7基因参与了幼体肌肉的发生过程。此外,进一步分析FcBMP-7基因在中国明对虾2种规格幼虾[(3.95±0.08) g和(1.55±0.12) g]的8种组织中的表达模式,结果显示,FcBMP-7基因具有组织表达广泛性,且两组间的大部分组织的表达量均存在显著差异(P<0.05)。试验结果表明,FcBMP-7基因可能作为调节因子参与了中国明对虾幼体肌肉发生和幼虾肌肉生长过程,研究结果可为进一步解析中国明对虾肌肉发生发育的分子机制提供重要的理论资料。In order to analyze the expression of bone morphogenetic protein-7(BMP-7)gene,a member of transforming growth factor beta(TGF-β)superfamily,in the muscle growth and development of Chinese shrimp Fenneropenaeus chinensis,the BMP-7(FcBMP-7)was cloned by through Rapid Amplification of cDNA Ends(RACE)technology.The full-length cDNA of FcBMP-7 was 2003 bp,containing 466 bp of 5′untranslated region(UTR),295 bp of 3′UTR and 1242 bp of open reading frame(ORF),encoding 413 amino acids.Using real-time PCR technology to analyze the expression pattern of FcBMP-7 in the muscle tissues of Chinese shrimp larvae at five different developmental stages(gastrulation stage,nauplius stage,zoea stage,mysis stage,and postlarval stage).The results showed that the gene was expressed in all stages of larval development,with the minimal expression in the gastrula stage,the significant increase in expression level from the nauplius stage(P<0.05),and the maximal expression level in the mysis stage;indicating that FcBMP-7 was involved in the development of shrimp muscle.Expression pattern of FcBMP-7 in 8 tissues of two sizes of Chinese shrimp revealed that FcBMP-7 had extensive tissue expression.There were significantly different in the expression levels in most tissues between the two groups(P<0.05),especially in the heart,intestine,muscle,periopod and pleopod(P<0.01).The findings suggested that FcBMP-7 gene played a certain role in the muscle growth and development of Chinese shrimp,which would lay a theoretical foundation for further analysis of the molecular mechanism of muscle development in Chinese shrimp.

关 键 词：中国明对虾 BMP-7基因 克隆 表达模式 

分 类 号：Q78[生物学—分子生物学]