用RAPD技术对利什曼原虫kDNA、nDNA的分析

目的 :用RAPD技术分析利什曼原虫的kDNA及nDNA。方法 :用 7种随机引物扩增L .infantum和L .donovaniGS2的kDNA和nDNA ,分析扩增结果并计算两虫种间kDNA和nDNA共享度F值。结果 :7种随机引物对两虫种的kDNA、nDNA均扩增出各自特有带型。L .infantum和L .donovaniGS2两虫种间kD NA、nDNA扩增片段的F值分别为 0 2 2 7和 0 2。结论 :kDNA和nDNA均可作为利什曼原虫的RAPD扩增模板 ,这 7种引物均可用于对L .infantum和L .donovaniGS2进行RAPD分析。该两虫种kDNA、nDNA的共享度较低 ,说明二者遗传距离较远。在进行RAPD分析时 ,提取全DNA (包括kDNA和nDNA)进行扩增即可

ANALYSIS OF kDNA AND nDNA OF LEISHMANIA BY RAPD

AIM:To analyze kDNA and nDNA of Leishmania spp by RAPD. METHODS:Both kinetoplast DNA(kDNA) and genomic DNA(nDNA) of L.infantum and L.donovani GS2 isolates were amplified with seven different random primers. The amplified fragments were analyzed and F values of kDNA and nDNA between the two Leishmania species were calculated. RESULTS:The seven primers yielded characteristic patterns for nDNA and kDNA between L.infantum and L.donovani GS2 isolates under the same experiment condition and significant differences were found between the 2 species. F values of amplified kDNA and nDNA fragments between L.infantum and L.donovani GS2 were 0^227 and 0^2 respectively,indicating far genetic distance between them. CONCLUSION:The seven primers may be used to analyze L.infantum and L.donovani GS2 isolates with kDNA and nDNA as template. kDNA may not be pecially extracted because it was shown to have no more advantage than nDNA.