耐碳青霉烯类鲍曼不动杆菌OXA-23-like基因的克隆和表达  

Cloning and Expression of OXA-23-like Gene from Carbapenem-resistant Acinetobacter baumannii

作　　者：范学财[1] 郭朝阳[1] 郭宇航[2] 张吉生[2] Fan Xuecai;Guo Chaoyang;Guo Yuhang;Zhang Jisheng(The First Affiliated Hospital of Jiamusi University,Jiamusi,154002;The Second Affiliated Hospital of Jiamusi University,Jiamusi,154007)

机构地区：[1]佳木斯大学附属第二医院,佳木斯154002 [2]佳木斯大学附属第一医院,佳木斯154007

出　　处：《基因组学与应用生物学》2020年第3期1331-1336,共6页Genomics and Applied Biology

基　　金：黑龙江省省属本科高校基本科研业务费科研项目资助课题(2017-KYYWF-0567);黑龙江省卫生计生委科研项目(2017-405)共同资助。

摘　　要：扩增耐碳青霉烯类鲍曼不动杆菌(Acinetobacter baumannii) OXA-23-like基因,并表达纯化该蛋白,为深入研究鲍曼不动杆菌亚单位蛋白疫苗提供理论基础。从60份样品中分离并扩增出OXA-23-like基因,构建于p GEX-6p-1表达载体中,用BL21表达宿主细胞诱导表达并纯化蛋白;免疫印迹试验(Western blotting)验证OXA-23-like蛋白保守性。结果显示,成功构建pGEX-6p-1-OXA-23-like质粒,表达并纯化蛋白;Western blotting实验表明临床菌株OXA-23-like蛋白表达阳性。OXA-23-like基因和蛋白表达保守性高,具有免疫原性,是鲍曼不动杆菌疫苗良好的抗原靶点。To amplify the OXA-23-like gene of carbapenem-resistant Acinetobacter baumannii and express and purify the protein, which lays a theoretical foundation for further study of Acinetobacter baumannii subunit protein vaccine. The OXA-23-like gene was isolated and amplified from 60 samples, constructed in pGEX-6 p-1 expression vector, and expressed and purified by BL21 expression host cells. Western blotting was used to verify OXA-23-like protein conservation. The pGEX-6 p-1-OXA-23-like plasmid was successfully constructed and expressed and purified. Western blotting analysis showed that the clinical strain OXA-23-like protein was positive. OXA-23-like gene and protein expression are highly conserved and immunogenic, which is a good antigen target for Acinetobacter baumannii vaccine.

关 键 词：耐碳青霉烯类鲍曼不动杆菌 原核表达 OXA-23-like 免疫印迹试验 

分 类 号：R978[医药卫生—药品]