[Construction of the subtracted cDNA libraries related to artemisinin-resistance of Plasmodium berghei].
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 250-2000 bp.


CONCLUSIONS
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.
Bei
Zhu-chun
ZC
Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.
Wang
Jing-yan
JY
D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology; D037621:Artemisinins / Q000494:pharmacology*; D018076:DNA, Complementary; D004351:Drug Resistance / Q000235:genetics*; D015698:Genomic Library; D008288:Malaria / Q000517:prevention & control*; D008297:Male; D051379:Mice; D009693:Nucleic Acid Hybridization / Q000379:methods; D010962:Plasmodium berghei / Q000187:drug effects* / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012717:Sesquiterpenes / Q000494:pharmacology*
D000890:Anti-Infective Agents; D037621:Artemisinins; D018076:DNA, Complementary; D012717:Sesquiterpenes; C031327:artemisinin