摘要
根据Gen Bank中公布的哈维氏弧菌qnr基因序列设计引物扩增哈维氏弧菌qnr序列,将其插入p ET-32a质粒,构建原核表达载体p ET32-qnr,并对诱导温度、时间、IPTG浓度等条件进行优化。结果表明,哈维氏弧菌qnr全长651 bp,编码216个氨基酸;重组蛋白优化条件为于28℃、IPTG浓度为0.05 mmol/L条件下诱导6 h。
The sequence of quinolone resistance gene ofVibrio harveyi was cloned based on theqnr of Vibrio harveyi published on GenBank and then inserted into the pET-32a vector to construct prokaryotic expression plasmid pET32-qnr.Then the expression conditions of temperature, induction time,and IPTG concentration of the pET32-qnr were optimized.The results showed that the completeopen reading frame (ORF) length ofqnrgene was 651bp,encoding a protein of 216 amino acids. The optimization conditions of inducible expression for Qnr protein wereinducingin 0.05 mmol/L IPTG for 6 hoursat 28℃.
出处
《广东海洋大学学报》
CAS
2016年第1期93-97,共5页
Journal of Guangdong Ocean University
基金
农业部行业专项
渔药使用风险评估及其控制技术研究与示范(201203085)
广东省教育厅高等学校高层次人才项目(谷胱甘肽及其合成酶系在哈氏弧菌耐药中的作用机制研究)
关键词
哈维氏弧菌
QNR基因
原核表达
优化
Vibrio harveyi
quinolone resistance gene
prokaryotic expression
optimization
