重组抗菌肽基因克隆及其Pichia pastoris表达工程菌的构建  

Cloning of antibaterial peptide gene and recombinanting in Pichia pastoris GSll5 strain

作　　者：彭梅[1] 尹娜[1] 谢天宏[1] 葛长勇[1] 张光明[1] 李鸿钧[1] 孙茂盛[1] 

机构地区：[1]中国医学科学院/北京协和医学院医学生物学研究所,云南昆明650118

出　　处：《云南大学学报（自然科学版）》2007年第S3期454-457,共4页Journal of Yunnan University(Natural Sciences Edition)

基　　金：云南省自然科学基金资助项目(2004C0029Q)

摘　　要：根据毕赤酵母对遗传密码的偏爱性,在不改动氨基酸序列的前提下,用化学法合成抗菌肽麻蝇素A的基因片段,合成片段拼接后,与α因子重组,重组基因再构建到表达载体pPIC9K上,得到受乙醇氧化酶1基因(AOX1)的启动子与转录终止区控制的酵母表达质粒,经鉴定分析,阳性重组子转化pastoris GS115宿主菌,经过表型筛选,成功获得了高效分泌表达麻蝇素A的重组毕赤酵母工程菌.A 120 bp DNA fragment encoding antibacterial peptide of sarcophaga peregrina A was designed based on the amino acid sequence of antibacterial peptide and the biased codon usage of pastoris. Eight oligonucleotides were chemically synthesized, linked and then recombinant with a- factor gene, recombinanted gene was cloned into yeast expression vector pPIC9K. After restriction enzyme analysis and DNA sequencing, the recombinanted gene of antibacterial peptide was transfected the Pichia pastoris GS115 strain. The positive clones screened by the phenotype were induced by methanol, The results showed that antibacterial peptide gene was expressed in Pichia pastoris successfully.

关 键 词：基因克隆 抗苗肽 PICHIA PASTORIS 

分 类 号：Q78[生物学—分子生物学]