肺炎衣原体重组的RNase HIIa和RNase HIIb的生化性质鉴定与比较  

Biochemical characterization and comparison of recombinant RNase HIIa and RNase HIIb from Chlamydia pneumoniae

作　　者：裴冬丽[1] 刘建华[2] 

机构地区：[1]商丘师范学院,河南商丘476000 [2]上海交通大学生命科学技术学院,上海200030

出　　处：《上海交通大学学报（医学版）》2008年第4期371-375,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(30170211)~~

摘　　要：目的对肺炎衣原体AR39的RNase HIIa和RNase HIIb(CpRNase HIIa和CpRNase HIIb)进行克隆、表达及生化性质鉴定与比较。方法按已知的CpRNase HIIs基因设计引物、扩增。用pET表达系统构建重组质粒,表达和纯化CpRNase HIIs。5′-32P标记底物RNA/DNA及含有核糖核苷酸的DNA,鉴定酶学性质。结果CpRNase HIIa和CpRNase HIIb均能切割RNA/DNA杂合体的寡聚核糖核苷酸链,产生3′羟基和5′磷酸基团的末端。但两者确实存在着不同的酶活性质。结论CpRNase HIIa和CpRNase HIIb在细胞内担负着不同的功能。Objective To clone and compare RNase HIIa and RNase HIIb of Chlamydia pneumoniae AR39（CpRNaseHIIa and CpRNaseHIIb）. Methods Genes of CpRNase HIIs were amplified with the designed primers.Then,the recombinant plasmids were constructed,and CpRNase HIIs were expressed and purified with pET expression system.The 5′-32P-labeled RNA/DNA substrate and DNA with oligoribonucleotides substrates were prepared to identify characterization of CpRNase HIIs. Results Ribonuclease H activity of both CpRNase HIIa and CpRNase HIIb could cleave oligoribonucleotides strand,generating break with 3′-OH and 5′-phosphate ends.The results showed that there was different biochemical characterization between them. Conclusion CpRNase HIIa and CpRNase HIIb have different functions in C.pneumoniae.

关 键 词：肺炎衣原体RNase HIIa 性质 

分 类 号：Q93[生物学—微生物学]