结核分枝杆菌对氨基水杨酸耐药相关基因的筛选及鉴定

Screening and identification of the PAS resistance related genes

目的筛选二线抗结核药物对氨基水杨酸(para-aminosalicylic acid,PAS)的耐药相关基因。方法构建H37Rv转座子文库,筛选PAS耐药克隆,随机引物PCR扩增转座子插入突变基因。在PAS耐药菌株中鉴定耐药相关基因的突变情况。结果构建了较完整的H37Rv转座子文库,并筛选出201个PAS耐药克隆,经测序找到4个可能的PAS耐药相关基因,分别是Rv1534、pknF、ndh、papA1。papA1基因检测到一个无义突变,为papA1基因339位C→T点突变。结论酰基转移酶基因papA1可能为PAS耐药相关基因。

Objective To screen the PAS （para-aminosalicylic acid） resistant-related genes. Methods The H37Rv transposon-insertion library was constructed. The PAS resistant clone was screened with random primer PCR, and mutant gene with transposon-insertion was identified. Furthermore, the mutants of the PAS resistant-related genes were examined in the PAS clinical resistant strain. Results The H37Rv transposon-insertion library was constructed successfully, and 4 probably PAS resistant-related genes （Rv1534,pknF.ndh.papA1） were identified. A nonsense mutation of papa 1 was found in the PAS clinical resistant strain. Conclusion papA 1 is probably the PAS resistant-related gene.