近平滑念珠菌ERG 11基因编码区的克隆及生物信息学分析

中国真菌学杂志　 2010, Vol. 5 　Issue (2): 92-96.

近平滑念珠菌ERG 11基因编码区的克隆及生物信息学分析

冯丽娟, 万喆, 王晓红, 李若瑜, 刘伟

北京大学第一医院皮肤性病科, 北京大学真菌与真菌病研究中心, 北京, 100034

收稿日期:2009-08-06 出版日期:2010-04-28 发布日期:2010-04-28
通讯作者: 刘伟,E-mail:liuw90@yahoo.com E-mail:liuw90@yahoo.com
作者简介:冯丽娟,女(汉族),硕士研究生在读.E-mail:flyingfeng2006@sina.com
基金资助:
教育部科学技术研究重点项目(107002)

Cloning and bioinformatic analysis of ERG11 gene in Candida parapsilosis

FENG Li-juan, WAN Zhe, WANG Xiao-hong, LI Ruo-yu, LIU Wei

Department of Dermatology, Peking University First Hospital, and Research Center for Medical Mycology, Peking University, Beijing 100034, China

Received:2009-08-06 Online:2010-04-28 Published:2010-04-28

摘要/Abstract

摘要： 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。

关键词: 近平滑念珠菌, ERG11基因, 克隆测序

Abstract: 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.

Key words: C.parapsilosis, ERG11 gene, cloning and sequencing

中图分类号:

R379.4

冯丽娟, 万喆, 王晓红, 李若瑜, 刘伟. 近平滑念珠菌ERG 11基因编码区的克隆及生物信息学分析[J]. 中国真菌学杂志, 2010, 5(2): 92-96.

FENG Li-juan, WAN Zhe, WANG Xiao-hong, LI Ruo-yu, LIU Wei. Cloning and bioinformatic analysis of ERG11 gene in Candida parapsilosis[J]. Chinese Journal of Mycology, 2010, 5(2): 92-96.

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