大劣按蚊感染约氏疟原虫defensins基因的克隆及生物信息学分析  

Cloning and Sequence Analysis of the Defensins from Anophele dirus infected P. yoelii

作　　者：张锡林[1] 王英[1] 陈继德[1] 宋蓓[1] 

机构地区：[1]第三军医大学基础部病原生物学教研室,重庆400038

出　　处：《成都医学院学报》2011年第2期94-98,共5页Journal of Chengdu Medical College

基　　金：国家自然科学基金项目(No.30800959;30872366)

摘　　要：目的为探讨大劣按蚊对约氏疟原虫侵入的免疫反应,克隆大劣按蚊的抗菌肽defensins基因并对基因序列进行生物信息学分析。方法分别提取感染疟原虫的大劣按蚊的蚊胃总RNA,根据冈比亚按蚊defensins基因设计、合成简并引物,进行RT-PCR扩增目的片段,TA克隆后测序并进行序列分析。结果用简并引物的RT-PCR扩增出约氏疟原虫感染的大劣按蚊的189bp目的片段,序列分析表明与冈比亚按蚊defensins有87%同源性,与白蚊伊蚊defensinsD有88%同源性,为大劣按蚊的defensins基因。结论成功克隆并分析了大劣按蚊抗菌肽defensins基因,为进一步的重组表达和研究按蚊抗菌肽的生物活性奠定基础。Objective Objective To research immune response of Anophele dirus and clone the defensins genes from Anophele dirus infected Plaasmodium yoelii and conduct sequence analysis.Methods Total RNA were extract from the gut of An.dirus infected P.yoelii,followed by RT-PCR with 1 pairs of degenerate primer designed and synthesized on the basis of reported defensins gene sequence of An.gambiae.The product of PCR were purfied and cloned into plasmid T-vector.The recombinant plasmid was sequenced and analyzed.Result The amplified products were about 189 bp from An.infected Plaasmodium yoelii by RT-PCR with degeneracy primer..Sequence analysis showed it had homology with 87% compared with the defensins sequence from An.gambiae and 88% compared with the defensins D of Aedes albopictus.Conclusions The defensins genes of An.dirus infected P.yoelii have been cloned successfully.This will lay the foundation for the research of expression of the recombinant defensins and its antibiotic activity.

关 键 词：大劣按蚊 defensins基因 克隆 序列分析 

分 类 号：Q785[生物学—分子生物学]