扬州大学学报（农业与生命科学版）2017，Vol.38Issue(1)：6-11,6.DOI:10.16872/j.cnki.1671-4652.2017.01.002
淋病奈瑟菌WHO-A株Cas蛋白的表达和多克隆抗体制备
Prokaryotic expression and polyclonal antibodies preparation of Cas proteins of Neisseria gonorrhoeae WHO-A strain
摘要
Abstract
To explore the structure and functions of CRISPR system in Neisseria gonorrhoeae,genomic DNA of N.gonorrhoeae WHO-A strain was extracted and used as PCR template to clone gonococcal CT and TM Cas genes that were used to prepare recombinant Cas proteins in Escherichia coli.The rCT and rTM expressed by pCold-TF vector were taken to immunize mice to prepare specific antibodies,while the titers were detected using pGEX-6p-1 expressed recombinant Cas proteins.SDS-PAGE analysis indicated that the recombinant antigens with the expected molecular weights were obtained.The titers of CT or TM specific antibodies in immune sera reached 1 ∶ 25 600 or 1 ∶ 12 800 detected by indirect ELISA.The purified recombinant Cas proteins and the specific antibodies developed in this study provided experimental materials for the detection of Cas proteins in gonococci and the studies of the mechanisms of gonococcal CRISPR systems.关键词
淋病奈瑟菌/Cas蛋白/原核表达/多克隆抗体Key words
Neisseria gonorrhoeae/cas protein/prokaryotic expression/polyclonal antibody分类
医药卫生引用本文复制引用
卞晓锐,徐桐桐,单彩龙,朱怡雯,焦红梅,阴银燕,孔桂美,李国才..淋病奈瑟菌WHO-A株Cas蛋白的表达和多克隆抗体制备[J].扬州大学学报（农业与生命科学版）,2017,38(1):6-11,6.基金项目
国家自然科学基金资助项目(81471906) （81471906）
江苏省高校自然科学基金资助项目(14KJB310025) （14KJB310025）
扬州大学科技创新团队培养对象资助项目(201609) （201609）
