【作者】 魏晓莉； 李国福； 李刚； 赵京花； 吴德昌； 时云林；

【Author】 WEIXiao-li,LI Gang, SHI Yun-lin, et al. Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071

【机构】 中国人民解放军军事医学科学院微生物流行病研究所疟疾室； 军事医学科学院放射医学研究所分子毒理室；

【摘要】 目的 研究药物敏感虫株伯氏疟原虫K173株及其在本芴醇压力下产生的本芴醇抗性株之间基因表达的差异状况,克隆抗性相关基因.方法 采用mRNA差异显示技术寻找相关基因,并用Northern杂交证实.结果 共采用24种引物组合进行mRNA差异显示,克隆到30余个差异表达基因片段,对其中8个进行了Northern杂交鉴定及测序.结果表明:与敏感株相比,克隆CB52在本芴醇抗性株中表达明显增多,而在氯喹抗性株中表达有所减少,说明克隆CB52确与本芴醇抗性相关,序列同源性检索表明CB52序列与恶性疟原虫环亲蛋白基因有微弱同源性.同时向GenBank登录8条新基因序列.结论 克隆CB52基因表达水平的改变与伯氏疟原虫本芴醇抗性相关,疟原虫对本芴醇的抗性机制在基因水平不同于氯喹抗性.更多还原

【Abstract】 Objective To isolate genes related to benflumetol resistance in the rodent malaria Plasmodium berghei K173 strata Methods The mRNA differential display technique was used to identify differential expressed genes between drug-sensitive Plasmodium berghei K173 strain and benflumetol-resistant strain selected under benflumetoi pressure. Northern blots were performed in order to determine the genes actually derived from maternal strains. Results PCR reactions were prepared respectively with twenty-four pairs of primers in each of strains, thirty-three differential displayed genes fragments were cloned and eight of them were sequenced. The eight sequences were novel ESTs and were submitted to GenBank. Homology searching revealed that clone CB52 was slightly homologous to Plasmodium falciparum cyclophilin gene. Hybridization confirmed that the gene fragment CB52 was upregulated in the benflumetol-resistant parasite strain, while downregulated in the chloroquine-resistant strain. Conclusion CB52 fragment was involved in the benflumetol resistance in plasmodium berghei; and the genetic mechanisms of drug-resistance to benflumetol and chloroquine were different.更多还原