新生隐球菌5种血清型的CAP64基因的序列分析  

Molecular analysis of CAP64 gene sequences from five serotypes of Cryptococcus neoformans

作　　者：邹先彪[1] 廖万清[2] 温海[2] 吴建华[2] 仇芸[2] 杨金水[3] 

机构地区：[1]解放军总医院第一附属医院皮肤科,北京100048 [2]上海第二军医大学长征医院皮肤科,200003 [3]复旦大学遗传所,上海200433

出　　处：《检验医学与临床》2010年第11期1025-1026,1029,共3页Laboratory Medicine and Clinic

基　　金：国家自然基金资助项目(NO.39770002)

摘　　要：目的多糖荚膜是新生隐球菌的重要的毒性因子,应用荚膜多糖抗原可以进行新生隐球菌的血清分型。方法该研究分析了5种血清型的CAP64基因的序列的相似性,作者扩增了CAP64基因一个约550bp的DNA序列,扩增产物经纯化后克隆到T载体上,转化至大肠杆菌JM109株中,通过聚合酶链反应(PCR)鉴定阳性克隆并测序。结果 5种血清型CAP64序列的比较显示其间的碱基差异为61～91bp,相似性为83.75%～88.03%,其核酸序列的差异是多位点的,主要在227～231bp之间;氨基酸的分析结果与此类似。结论 5种血清型的CAP64基因存在着一定的差异。Objective The extracellular polysaccharide capsule produced by C.neoformans is essential for its pathogenicicty.The serotyping of C.neoformans has been performed immunologically with antisera against the mucopolysaccharide capsule component of the yeast.Methods The nucleotide sequence of CAP64 genes from five serotypes of C.neoformans were analyzed for their similarity.Approximately 550 bp genomic DNA fragments of the CAP64 gene were amplified from each isolate by PCR,the PCR products from each isolate were purified and cloned into pGEM-T vectors respectively,after transfecting JM109 strain,the recombinants were screened and identified by PCR and sequenced.Results A comparison of CAP64 sequences among the five serotypes indicated the presence of the ciding regions varing in size from 67 to 91 bp,and the similarites were from 83.75% to 88.03%.Nucleotide sequence analysis of CAP64 gene fragments indicated that distinct the variant sites of nucleotides appear aporadically,sequences and mostly between 227 bp and 231 bp.The results of amino acid sequences were similar to the nucleotide sequence data.Conclusion There is certain difference in CAP64 sequences among the five serotypes of Cryptococcus neoformeans.

关 键 词：新生隐球菌 CAP64基因 序列 

分 类 号：R378.1[医药卫生—病原生物学]