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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
0
(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pPICZαA,构建重组表达质粒pPICZαA-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/pPICZαA-NPY,用终浓度为1%的甲醇诱导表达阳性转化菌,用Tricine-SDS-PAGE和Western Blot检测表达蛋白。结果成功构建表达载体pPICZαA-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:    毕赤酵母  神经肽Y  基因表达
DOI:10.13610/j.cnki.1672-352x.20170208.025
投稿时间:2016-07-04
基金项目:国家自然科学基金项目(31572404)资助。
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 pPICZαA vector to obtain recombinant pPICZαA-NPY, and then the recombinant plasmid was transfected into Pichia pastoris GS115 by electrotransformation. The positive recombinant strains were induced to express the recombinant protein with 1% methanol. Tricine-SDS-PAGE and Western blot were used to detect the recombinant protein. The results showed that the recombinant NPY protein could be secreted 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

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