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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 :采用 2 4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 ,而在氯喹抗性株中的表达有所减少 ,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。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的产生相关 ,疟原虫对本芴醇的抗性机制在基因水平不同于氯喹抗性
OBJECTIVE: To study the difference of gene expression between drug-resistant strains of Plasmodium berghei K173 and the strain of lylroflumethrol resistant to lumefantrine resistance, and cloned resistance genes. Methods: The mRNA differential display technique was used to find the related genes and confirmed by Northern blotting. Results: 24 pairs of primers were used for mRNA differential display. More than 30 differentially expressed genes were cloned and 8 of them were identified by Northern hybridization and sequencing. The results showed that, compared with the susceptible strain, the expression of cloned CB52 was significantly increased in the benflumetol resistant strains, while the expression decreased in the chloroquine resistant strains, indicating that the clone CB52 was indeed related to the resistance to the benflumetol. The sequences Source-based searches revealed a weak homology between the CB52 sequence and the P. falciparum cyclodextrin gene. Eight new gene sequences were registered with GenBank. Conclusion: The change of CB52 gene expression level is related to the resistance of P. berghei to lumefantrine. The resistance mechanism of P. falciparum to lumefantrine is different from that of chloroquine resistance at gene level