乳酸乳球菌内源隐蔽性质粒序列分析及食品级克隆载体的构建  

Sequence Analysis of Lactococcus lactis Cryptic Plasmid and Development of a Food-grade Cloning Vector

作　　者：范丽平[1] 张江[1] 霍贵成[2] 

机构地区：[1]上海农林职业技术学院农业生物与生态技术系,上海201699 [2]乳品科学教育部重点实验室(东北农业大学),哈尔滨150030

出　　处：《中国食品学报》2015年第5期182-188,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家"863"项目(No.2002AA248041)

摘　　要：为构建乳酸菌食品级载体,对乳酸乳球菌内源隐蔽性质粒p KL001进行全序列测定、序列分析及PCR扩增,获得了该质粒的复制子。以Southern杂交对质粒p KL001复制模式进行分析,结果显示p KL001为θ型复制。将该复制子与镉抗性功能片段连接,构建了食品级克隆载体p KF168,其在受体菌株KLDS4.0319-5中可稳定存在。In order to construct food-grade cloning vector, the complete nucleotide sequence of the cryptic plasmid, designated pKL001 has been determined. After sequence analysis, the fragment of pKL001 with the function of replica- tion was amplified. The result of Southern blot indicates that pKL001 has theta-type mechanism of replication. The food grade-cloning vector pKF168 was constructed using the replicon of pKL001 and the cadmium resistance select marker. It was stable in KLDS4.0319-5 for at least 48 generations without selection pressure.

关 键 词：乳酸乳球菌 隐蔽性质粒 复制子 克隆载体 

分 类 号：TS201.3[轻工技术与工程—食品科学]