Construction of the Subtracted cDNA Libraries Related to Artemisinin-resistance of Plasmodium berghei
- VernacularTitle:伯氏疟原虫青蒿素抗性相关的消减cDNA文库构建
- Author:
Zhuchun BEI
;
Jingyan WANG
- Publication Type:Journal Article
- Keywords:
Plasmodium berghei;
Artemisinin;
Drug resistance;
Polymerase Chain Reaction;
Gene Libraries
- From:
Chinese Journal of Parasitology and Parasitic Diseases
1987;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the subtracted cDNA libraries related to artemisinin-resistance of Plasmodium berghei using suppression subtractive hybridization PCR (SSH PCR). Methods Total RNA was extracted from the artemisinin-sensitive (NS) and artemisinin-resistant (AR) strains of Plasmodium berghei K173. The cDNA synthesis followed the protocol of super SMART cDNA synthesis kit. Taking the NS as driver, AR as tester and reverse,two subtractions were performed by SSH PCR. Enriched different expressed cDNA was cloned into pMD18-T vector to construct subtractive libraries. Results The subtracted cDNA libraries of NS-AR and AR-NS contained 395 and 506 positive clones respectively. The PCR results of 108 clones picked randomly from each library showed 100 and 104 positive inserts contained in the plasmids respectively, and distributing in 250-2 000 bp. Conclusion The successful construction of the subtracted cDNA libraries related to artemisinin-resistance of P. berghei enable us to identify the different expressed genes involved in the resistance mechanism.
