鸡神经肽Y(NPY)基因在毕赤酵母中的表达.PDFVIP

安徽农业大学学报, 2017, 44(1): 40-43 Journal of Anhui Agricultural University [DOI] 10.13610/ki.1672-352x025 网络出版时间：2017/2/8 10:28:00 [URL] /kcms/detail/34.1162.S1028.050.html 鸡神经肽 Y （NPY ）基因在毕赤酵母中的表达 * 张 鑫，程军军，姜勋平，刘桂琼 (华中农业大学动物科技学院，武汉 430070) 摘 要：为通过毕赤酵母的表达获得鸡NPY 蛋白，根据 GenBank 上的鸡 NPY 基因序列，结合毕赤酵母密码子 的偏好性合成编码鸡NPY 蛋白的基因，将其插入真核表达载体pPICZ αA ，构建重组表达质粒pPICZ αA-NPY ，并 将重组质粒电转入毕赤酵母 GS115，获得重组毕赤酵母菌 GS115/pPICZ αA-NPY ，用终浓度为 1%的甲醇诱导表达 阳性转化菌，用Tricine-SDS和 Western Blot 检测表达蛋白。结果成功构建表达载体pPICZ αA-NPY ，转化后 的重组酵母菌成功分泌鸡NPY 蛋白，获得分子量约为 35 kDa 的鸡NPY 蛋白。 关键词：鸡；毕赤酵母；神经肽 Y ；基因表达 中图分类号：S831 文献标识码：A 文章编号：1672352X (2017)01004004 Expression of chicken NPY gene in Pichia pastoris ZHANG Xin, CHENG Junjun, JIANG Xunping, LIU Guiqiong (College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070) Abstract: The aim of this study was to obtain chicken neuropeptide Y（NPY ）protein. The DNA sequence of the chicken NPY gene from GenBank was modified based on the codon preference of Pichia pastoris . The modified DNA sequence was synthesized and inserted into pPICZ αA vector to obtain recombinant pPICZ αA-NPY , and then the recombinant plasmid was transfected into Pichia pastoris GS115 by electrotransformation. The positive recom- binant strains were induced to express the recombinant protein with 1% methanol. Tricine-SDSand Western blot were used to detect the recombinant protein. The results showed that the recombinant NPY protein could be se- creted by the positive recombinant strains and the molecular weight of NPY protein was approximately 35 kDa. Key words: chicken; Pichia pastoris ; neuropeptide Y; gene expression 神经肽 Y （NPY ）是一个由36 个氨基酸构成的