近平滑念珠菌ERG 11基因编码区的克隆及生物信息学分析

目的克隆、测序近平滑念珠菌ERG11基因的编码区序列并进行生物信息学分析。方法运用生物信息学的方法 ,通过与白念珠菌ERG11基因碱基序列同源性比对,在近平滑念珠菌基因组(www.sanger.ac.uk/sequencing/Candida/pa-rapsilosis/)中寻找可能的ERG11基因序列(CpERG11),并据此序列设计引物,经PCR扩增近平滑念珠菌标准株(ATCC22019)的ERG11基因片段,产物经电泳、纯化、克隆到质粒prG-AMAI-NotI中,转染DH10B大肠杆菌细胞,并酶切鉴定筛选阳性克隆测序分析。结果近平滑念珠菌ERG11编码区由1569个碱基组成,编码一段含522个氨基酸的多肽。近平滑念珠菌ERG11的编码区序列与白念珠菌、热带念珠菌、光滑念珠菌、酿酒酵母菌ERG11基因的同源性分别为74%、75%、65%、64%。该近平滑念珠菌ERG11的编码区为唑类药物作用靶酶基因。结论成功克隆、测序、并生物信息学分析近平滑念珠菌ERG11基因的编码区序列,为进一步的功能研究奠定基础。

Cloning and bioinformatic analysis of ERG11 gene in Candida parapsilosis

Objective To clone and analyze the coding region of ERG11 gene of Candida parapsilosis by bioinformatics.Methods The C.parapsilosis genome was compared to find the possible ERG11 gene sequence of Candida albicans by bioinformatics.The coding region of ERG11 gene of C.parapsilosis was amplified by primers designed based on the found sequence.The purified PCR product was then cloned into plasmid prG-AMAI-NotI and the recombinated plasmid was further transformed into E.coli DH10B for identification.Results The size of the open reading frame （ORF） of ERG11 gene from C.parapsilosis was 1 569 bp,encoding a peptide with 522 amino acid.The nucleotide sequence homelogy of ERG11 gene from C.parapsilosis was 74%,75%,65%,64% with that from C.albicans,Candida tropicalis,Candida glabrata,and Saccharomyces cerevisiae respectively.The ORF of C.parapsilosis ERG11 gene obtained in this study encoded the target enzyme for azoles antifungal drugs.Conclusions The ORF of ERG11 gene from C.parapsilosis was successfully cloned,sequenced and analyzed by bioinformatics,which provided the basis for further study on its function.