摘要
目的克隆和研究耐药性淋球菌相关基因.方法从耐药淋球菌株RSM292C4和淋球菌标准参考菌株WHO-A细胞中提取DNA,经Rsa Ⅰ酶切后,应用抑制性消减杂交技术构建耐药性淋球菌差异DNA消减文库并进行初步筛选.结果耐药淋球菌差异DNA消减文库经扩增获得约2 500个白色克隆,随机挑取96个白色克隆,进行PCR扩增鉴定,发现克隆中插有100～600 bp大小的片段.分别以RsaI酶切样本和参照DNA为探针,与随机挑取的96个白色菌落质粒作点杂交,有5个克隆仅能与样本DNA探针杂交,而不能与参照DNA探针杂交,表明它们为RSM292C4基因组DNA所特有,可能代表某些淋球菌新基因.对这5个克隆插入的DNA片段进行测序,经送基因库和淋球菌基因组部分测序基因库检索分析,发现5个片段均为未知新序列.结论淋球菌耐药菌株DNA消减文库的构建为进一步筛选、克隆淋球菌耐药的新基因奠定了基础.
Objective To clone and study the antimicrobial-resistant gene in Neisseria gonorrhoeae.Methods The gene library,which contains the differential genes of antimicrobial resistant strains and stan-dard reference strains of Neisseria gonorrhoeae,was constructed using a technique known as suppression subtractive hybridization(SSH).Then the antimicrobial resistance associated genes were cloned and ana-lyzed.Results Subtractive gene library in antimicrobial-resistant Neisseria gonorrhoeae was successfully constructed,which contains2500positive clones.Sequence analysis was performed on5clones.The se-quences of these five clones were unknown previously.Conclusions The subtractive DNA library is succes-sively constructed which may provide an important clue for studying the mechanism of antimicrobial resis-tance in Neisseria gonorrhoeae.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2003年第5期264-266,共3页
Chinese Journal of Dermatology
关键词
耐药淋球菌
抑制性消减杂交
基因文库
药物耐受
DNA序列分析
Neisseria gonorrhoeae
Drug resistance,bacterial
Suppression subtractive hybridiza-tion
Sequence analysis,DNA
Gene library
