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当前位置: 首页 > 文章 > 绵羊肌肉生长抑制素(MSTN)双价纳米抗体构建与表达 黑龙江畜牧兽医 2016 (17) 42-45
Position: Home > Articles > Construction and expression of bivalent nanobodies of sheep myostatin(MSTN) Heilongjiang Animal Science and Veterinary Medicine 2016 (17) 42-45

绵羊肌肉生长抑制素(MSTN)双价纳米抗体构建与表达

作  者：
吴鹏;黄美玲;李尤简;张勋;梁田
单  位：
石河子大学医学院;石河子大学动物科技学院;石河子大学动物科技学院新疆地方与民族髙发病教育部重点实验室;石河子大学
关键词：
肌肉生长抑制素(MSTN);纳米抗体;噬菌体文库;原核表达;Western-blot;蛋白纯化
摘  要：
为了研究如何提高纳米抗体颉颃肌肉生长抑制素(MSTN)作用,试验采用构建绵羊MSTN纳米抗体双价表达载体的方法,将免疫双峰驼构建的MSTN特异性纳米抗体噬菌体文库筛选得到的特异性抗MSTN纳米抗体,扩增目的序列,利用linker进行链接,将序列构建到原核表达载体p ET-30a,转化进BL21(DE3),经过SDS-PAGE电泳以及Western-blot检测,利用Ni柱纯化双价抗MSTN蛋白纳米抗体。结果表明:经过诱导的表达菌,SDS-PAGE验证以及Western-blot检测,条带清晰。说明成功构建了颉颃MSTN的双价纳米抗体原核表达系统;通过纳米抗体技术制备MSTN的颉颃物,阻断其分子作用通路,可有效地促进绵羊肌肉的生长和发育。
译  名：
Construction and expression of bivalent nanobodies of sheep myostatin(MSTN)
作  者：
WU Peng;HUANG Meiling;LI Youjian;ZHANG Xun;LIANG Tian;XIAO Yuanyuan;CHEN Xinkai;CHEN Chuangfu;SHENG Jinliang;College of Animal Science and Technology,Shihezi University;Key Laboratory of Ministry of Education for Xinjiang Endemic and Ethnic Diseases;
关键词：
myostatin(MSTN);;nanobody;;phage library;;prokaryotic expression;;Western-blot;;protein purification
摘  要：
To study how to improve the role of antagonistic nanobody myostatin( MSTN),MSTN nano- specific antibody phage libraries constructed by the bactrian camel immunization were screened to obtain specific anti- MSTN nanobodies using the method of constructing bivalent nanobody expression vector of sheep MSTN. The target sequences were amplified,and then were linked using linker to construct a prokaryotic expression vector p ET- 30 a. The expression vector was transformed into BL21( DE3),while SDS- PAGE electrophoresis and Western- blot detection were carried out. Purified by Ni bivalent anti- MSTN protein nanobodies were purified using Ni coulumn. The results showed that after induced expression of bacteria,SDS- PAGE and Western- blot strips were clear. The result indicates that the prokaryotic expression system of antagonistic MSTN bivalent nanobodies is successfully constructed. MSTN antagonists prepared by nanobody technique,blocks the molecular pathway,which can effectively promote muscle growth and development of sheep.

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