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机构地区:[1]安徽理工大学医学院病原生物学教研室,安徽淮南232001 [2]第三军医大学基础部病原生物学教研室
出 处:《中国病原生物学杂志》2006年第2期102-105,共4页Journal of Pathogen Biology
基 金:国家自然科学基金资助项目(No.30400363)。
摘 要:目的 比较分析对约氏疟原虫易感的斯氏按蚊和不易感的大劣按蚊的基因组DNA的多态性,探讨媒介按蚊与疟原虫遗传基因间的相互关系。方法 设计5条随机引物,应用随机扩增多态性DNA(random amplified polymorphic DNA,RAPD)技术对斯氏按蚊、大劣按蚊成蚊(未感染蚊和感染蚊)及其幼虫的基因组DNA进行随机扩增,扩增产物经琼脂糖凝胶电泳后分析。结果 5条随机引物对2种按蚊扩增的条带数不同,其中P5对斯氏按蚊扩增的效果较好,共16~17条带型,长度为200~1800bp;P4对大劣按蚊扩增效果较好,出现17条带型,长度为180~1500bp;P1对大劣按蚊无扩增产物。用P3对2种按蚊扩增,感染前后的扩增产物电泳条带数相同,但亮度不同;成蚊与幼虫带型数量不同,斯氏按蚊成虫与幼虫有3条相同带型(400、650、850bp),大劣按蚊成虫与幼虫有4条相同带型(350、500、750、800bp);大劣和斯氏按蚊成蚊带型分别为10和13条,其中相同带型6条(300、350、400、550、600、800bp)。结论 斯氏按蚊和大劣按蚊之间存在基因差异。同种未感染蚊和感染蚊基因带型仅存在亮度差异,而幼虫与成虫的基因差异较大。Objective To analyze the genomic DNA polymorphism of Anopheles dirus and Anopheles stephensi, so as to explore the relationship of the genomic DNA between the Anopheles and Plasmodium. Methods The genomic DNA extracted from the different stages of An. dirus and An. stephensi were amplified by random amplified polymorphic DNA (RAPD) with 5 random primers. And then the amplified products were carried on agarose gel electrophoresis. Results The genomic DNA of An. dirus and An. stephensi were successfully amplified by RAPD with 5 different primers. The primer P5 was better than others to An. stephensi, from which 16-17 bands between 200-1 800 bp were detected. The primer P4 was better than others to An. dirus, from which 17 bands between 180-1 500 bp were detected. No bands were obtained in electrophoresis of amplified product of An. dirus with the primer P1. An. dirus and An. stephensi were then amplified with primer P3 and P4, only a little difference in brightness between the normal and infected mosquito was discovered, no difference of bands quantity between the adult mosquito and larva. For example, with primer P3, 3 similar bands(400, 650, 850 bp) were detected in both the adult An. stephensi and larva, 4 similar bands(350, 500, 750, 800 bp)were detected in both the adult An. dirus and larva, 6 similar bands (300, 350, 400, 550, 600, 800 bp) were detected in both An. dirus and An. stephensi, but 7 different bands were also detected. Conclusion Genic variance was great between An. dirus and An. stephensi, only a little difference between the normal and the infected mosquitoes in brightness of genic bands, but there was great difference between adult and larva of two Anopheles.
分 类 号:R384.1[医药卫生—医学寄生虫学]
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