摘要
目的构建流产布鲁氏菌omp25基因毕赤酵母分泌型表达载体。方法根据目的基因序列分析结果和毕赤酵母对密码子的优先选择性,选择了抗原性较强的长471bp的基因片段进行克隆和构建流产布鲁氏菌omp25基因毕赤酵母分泌型表达载体,并予鉴定。结果鉴定结果表明目的基因片段与发表的序列完全一致并正确地插入到酵母表达载体pPIC9Kα因子分泌信号肽下游。结论成功地构建了流产布鲁氏菌omp25基因的毕赤酵母表达载体pPIC9K-omp25。
According to the results of sequence analysis on target gene and the preceding selectivity of Pichia. pastoris to codons, a gene fragment of 471 bp in length and with high antigenicity was cloned and amplified by PCR in order to construct the P. pastoris secretory expression vector pPIC-9K-omp25 containing the omp25 gene in Brucella abortus. The rccombinarnt P. pastoris was then identified. The identification result of this expression vector indicated the complete identity in sequence of the cloned omp25 gene of Br. abortus with those published, and that the target gene fragment could be accurately inserted into downsteam of α-factor selection signal peptide of P. pastoris exoression vector pPIC-gK （by alcohol oxidase AOX1 promoter）. It is concluded the P. pastons expression vector containing omp25 gene of Br. abortus was successfully constructed.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2007年第7期693-695,共3页
Chinese Journal of Zoonoses
基金
国家奶业重大专项(2002BA518A04
2006BAD04A05)
