重叠延伸PCR法扩增EB病毒BZLF1N-BLRF2融合基因及生物信息学分析  

Construction of Epstein-barr Virus BZLF1N-BLRF2 Gene by Overlap Extension PCR and Bioinformatics Analysis

作　　者：张毅[1,2] 周淑艳[1] 陈锐[1,2] 孙业红[1] 李体远[1] 

机构地区：[1]暨南大学附属第二医院深圳市人民医院临床医学研究中心,深圳518020 [2]暨南大学药学院,广州510632

出　　处：《生物技术通报》2012年第3期173-178,共6页Biotechnology Bulletin

基　　金：暨南大学科研培育与创新基金研究项目(216113103);深圳市卫生局重点课题(200608)

摘　　要：利用重叠延伸PCR技术扩增EB病毒BZLF1N-BLRF2融合基因,构建原核表达载体pGEX-4T-1-BZLF1N-BLRF2,并进行了蛋白的诱导表达。通过序列测定和ORF软件分析,该融合基因含有1个1 005 bp的ORF,编码335个氨基酸。应用生物信息学方法,对该融合基因编码的蛋白ZtaN-p23从氨基酸组成、理化性质、信号肽、疏水性/亲水性、二级结构、亚细胞定位、功能域和高级结构等方面进行了预测和分析。结果表明,该蛋白的预测分子量为46.2 kD,理论等电点约为8.97,是亲水性蛋白,推测它定位于细胞质中。序列分析表明,该蛋白可能具有信号转导、转录调控、免疫应答等功能。预测的二级结构及三级结构都表明该蛋白含有较多的不规则卷曲和α-螺旋,三级结构上呈对称形状。Epstein-Barr virus BZLF1N gene and BLRF2 gene were amplified and linked by overlap extension PCR method.The prokaryotic expression vector pGEX-4T-1-BZLF1N-BLRF2 was constructed,which expressed in E.coli.The fusion gene contained an ORF with the longest length of 1 005 bp,and encoding 335 amino acids.The physicochemical characteristics,structures and functions of fusion protein ZtaN-p23 which encoded by BZLF1N-BLRF2 were predicted and analyzed with software tools and database of bioinformatics.The results showed that the molecular weight and pI of deduced protein is 46.2 kD and 8.97 as well as it has not signal peptide,which is a hydrophilic protein locating in cytoplasm.The sequencing analysis displayed that the protein holds the possibility of signal transducer,transcription regulation and Immune response.The predicted second and tertiary structure indicates that ZtaN-p23 contains more random coil and helix and a small quantity of extended strand.

关 键 词：重叠延伸 PCR EB病毒 BZLF1N基因 BLRF2基因 生物信息学 

分 类 号：R440[医药卫生—诊断学]