%0 Journal Article
%T Construction of a food-grade secretion expression vector and use it for reporter protein expression in Lactococcus lactis<br>食品级分泌表达载体的构建及报告蛋白在乳酸乳球菌中的表达
%A Qiangzheng Sun
%A Yanwen Xiong
%A Changyun Ye
%A Jianguo Xu
%A <br>孙强正
%A 熊衍文
%A 叶长芸
%A 徐建国
%J 微生物学报
%D 2008
%I 
%X We constructed a food-grade secretion expression vector and used it for reporter protein expression in live delivery vehicle L lactisMBP71. The p32 fragment, which containing the stronger p32 promoter,,was amplified by polymerase chain reaction(PCR) with the plasmid pMG36e as template. After being purified, the p32 fragment was ligated with SPusp45 fragment amplified from genomic DNA of L lactis MG1363. The fusion fragment p32-SPusp45 was inserted into the food-grade vector pSH91 to construct a secretion expression vector, pSQ. The coding sequence of NucA (nucA) was also amplified from Staphylococcus aureus chromosome and inserted into pSQ under the control of p32 promoter to construct a recombinant plasmid pSQ-nucA. Nuclease plate activity assay and zymograme assay demonstrate that NucA was secretion expressed from L lactis harboring the recombinant plasmid pSQ- nucA, and the quantity of NucA secreted into supernamant was about ten times more than which in cell lysate. Results also indicate that expression efficiency of L lactis/pSQ-nucA was higher than that of L lactis/pSQZ-nucA, constructed by us earlier.
%K Lactococcus lactis
%K p32 promoter
%K food-grade vector
%K secretion expression
%K reporter protein<br>乳酸乳球菌
%K p32启动子
%K 食品级载体
%K 分泌性表达
%K 报告蛋白
%K 食品级
%K 表达载体的构建
%K 蛋白
%K 乳酸乳球菌
%K Lactococcus
%K lactis
%K protein
%K expression
%K expression
%K vector
%K secretion
%K 疫苗
%K 胞内
%K 分泌性表达
%K 发现
%K 结果
%K 比较
%K 表达能力
%K 表达系统
%K 表达量
%K 表达形式
%K 检测
%K 酶谱法
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=561F79F1A238AD5B180C9CB0EBE9A2FF&yid=67289AFF6305E306&vid=B6DA1AC076E37400&iid=38B194292C032A66&sid=2AC7DCCBBC26ECF8&eid=88D36036CFF69B3C&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=18