摘要
为提高重组乳酸菌NICE诱导表达系统表达外源蛋白的能力,研究选取Nisin加入量、OD值、诱导温度和诱导时间这4个主要影响表达的因素,每个因素设置3个水平,设计4因素3水平(43)正交试验。通过优化Nisin加入量、OD值、诱导温度和诱导时间,从而确定重组乳酸菌表达外源蛋白最佳条件。结果显示:当Nisin的量为1 ng/mL、OD值为0.3、诱导温度为30℃、诱导时间为7 h时,其重组乳酸菌表达外源蛋白的量最高。研究结果为由Nisin控制表达基因系统的重组乳酸乳球菌疫苗开发奠定了基础。
In order to improve the ability of recombinant lactic acid bacteria NICE induced expression system to express foreign proteins, this study selected three levels from each factor based on the four main factors affecting the expression of Nisin supplementation, OD value, induction temperature and induction time. The orthogonal experimental table was designed for 4 factors and 3 levels. By optimizing the three levels of Nisin supplementation, OD value,induction temperature and induction time, the optimal conditions for the expression of exogenous proteins in recombinant lactic acid bacteria were determined. The results showed that when the amount of Nisin was 1 ng/mL, OD value was0.3, the induction temperature was 30 ℃ and the induction time was 7 h, the recombinant lactic acid bacteria expressed the highest amount of foreign protein. This study laid the foundation for the vaccine development of recombinant Lactococcus lactis with the gene expression system controled by Nisin.
作者
张园
王文骞
宋雨心
王婷婷
刘琳琳
崔红玉
何高明
ZHANG Yuan;WANG Wenqian;SONG Yuxin;WANG Tingting;LIU Linlin;CUI Hongyu;HE Gaoming(College of Animal Science and Technology,Shihezi University,Shihezi,Xinjiang 832003;State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin,Heilongjiang 150069)
出处
《中国家禽》
北大核心
2019年第5期25-28,共4页
China Poultry
基金
国家自然科学基金面上项日(31570930)
国家重点研发计划项目(2017YFD0500802-05)
关键词
重组乳酸乳球菌
正交试验法
诱导表达
recombinant Lactococcus lactis
orthogonal experiment
induced expression
作者简介
张园(1994-),女,硕士研究生,研究方向为乳酸菌免疫调节研究,E-mail:18089218836@163.com;通讯作者:崔红玉(1970-),男,副研究员,博士,主要从事乳酸菌免疫调节研究,E-mail:cuihongyu@caas.cn;通讯作者:何高明(1967-),男,博士,教授,主要从事动物营养代谢病防治研究,E-mail:hgms@shzu.eu.cn。
