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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (17): 101-104.doi: 10.11924/j.issn.1000-6850.2011-3555

所属专题： 生物技术

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

TAT-NPY融合蛋白的表达和生物活性检测

费东亮 张才 刘国文 王哲   

  • 收稿日期:2011-11-29 修回日期:2011-12-15 出版日期:2012-06-15 发布日期:2012-06-15

Expression and Bioactive of TAT-NPY Fusion Protein

  • Received:2011-11-29 Revised:2011-12-15 Online:2012-06-15 Published:2012-06-15

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摘要：

摘 要：为了研究含有跨膜结构域TAT基因的重组奶牛神经肽Y融合蛋白的生物学活性。利用基因工程技术，在牛神经肽Y基因序列中插入TAT蛋白基因序列，构建原核表达载体pGEX-3X-NPY-PTD，导入到BL21宿主菌中，进行表达,对表达产物进行鉴定、表达条件优化和纯化。并且经过活性检测,融合蛋白对鸡胚血管生成的促进作用,表明含有PTD结构的重组奶牛神经肽Y具有生物功能活性，为动物试验奠定了基础。为了获得重组TAT-NPY融合蛋白，并研究其生物学活性。利用基因工程技术，从犊牛下丘脑组织中克隆得到神经肽Y基因序列，通过基因比对符合率为100%。同时，在牛神经肽Y基因序列中插入TAT蛋白基因序列，构建原核表达载体pGEX-3X-NPY-PTD，导入到BL21宿主菌中，进行原核表达，并对表达产物进行鉴定、表达条件优化和纯化。并且经过活性检测，融合蛋白对鸡胚血管生成起促进作用。表明含有PTD结构的重组奶牛神经肽Y具有生物功能活性，为下一步的研究提供了物质基础。

关键词: 区划, 区划

Abstract:

In order to obtain the recombinant protein NPY containning TAT protein transduction domain and study the bioactive of the recombinant protein. NPY gene was cloned from the Calf Hypothalamus tissue by the molecular biology techniques. Coincidence rate was 100% by genetic test. TAT-protein gene was inserted into bovine NPY gene sequences and expression vector pGEX-3X-NPY-TAT was constructed, then the recombinant plasmids of pGEX-3X-NPY-PTD was induced in E. coli DE3 by addition of IPTG. The expressed fusion protein was identified, It was optimized and the bioactive test was carried out by CAM. The results showed that the bovine NPY gene of TAT protein transduction domain was expressed, and the expressed fusion protein had biological activity. Conclusion the study provided important base for the animal experiment.