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当前位置: 首页 > 文章 > 牛源犬新孢子虫NcSRS2-NcGRA7融合基因的克隆与生物信息学分析 畜牧与兽医 2013,45 (1) 25-28
Position: Home > Articles > Cloning and bioinformatics analysis of NcSRS2-NcGRA7 fusion gene of bovine Neospora caninum Animal Husbandry & Veterinary Medicine 2013,45 (1) 25-28

牛源犬新孢子虫NcSRS2-NcGRA7融合基因的克隆与生物信息学分析

作  者：
贾立军;张守发;谷振甲
单  位：
吉林省安图县畜牧兽医局;延边大学农学院动物医学系
关键词：
牛;犬新孢子虫;NcSRS2-NcGRA7;克隆;生物信息学
摘  要：
为了解牛源犬新孢子虫NcSRS2-NcGRA7融合基因的生物学特性,本试验提取牛源犬新孢子虫基因组DNA,应用PCR技术扩增犬新孢子虫表面蛋白基因NcSRS2和致密颗粒蛋白基因NcGRA7,SOE-PCR技术拼接NcSRS2和NcGRA7基因,构建NcSRS2-NcGRA7融合基因重组克隆质粒,并进行PCR鉴定、双酶切鉴定及生物信息学分析。结果,NcSRS2基因扩增片段大小为1 061 bp,NcGRA7基因扩增片段大小为364 bp,NcSRS2-NcGRA7融合基因扩增片段大小为1 482 bp;获得重组克隆质粒pMD18-NcSRS2-NcGRA7经PCR鉴定、双酶切鉴定正确;测序分析表明,与Gen-Bank中已发表的美国株犬新孢子虫(AF061249、AF176649)核苷酸序列同源性为99%;经DNAman等软件分析,预测NcSRS2-NcGRA7融合蛋白抗原指数较高,融合蛋白二级结构以α-螺旋和β-折叠为主,三级结构中2种蛋白独立折叠,并借助Linker互相连接,功能互不影响。本试验为牛源犬新孢子虫NcSRS2-NcGRA7融合蛋白的免疫学研究奠定了基础。
译  名：
Cloning and bioinformatics analysis of NcSRS2-NcGRA7 fusion gene of bovine Neospora caninum
作  者：
JIA Li-jun1,ZHANG Shou-fa1,GU Zhen-jia2(1.Department of Veterinary Medicine,Agriculture College of Yanbian University,Yanji 133002,China; 2.Bureau of Animal Husbandry and Veterinary Medicine,Antu 133600,China)
关键词：
bovine;Neospora caninum;NcSRS2-NcGRA7;cloning;bioinformatics analysis
摘  要：
In order to understand the biological characteristic of NcSRS2-NcGRA7 fusion gene of bovine Neospora Caninum,genomic DNA of N.caninum was extracted.The surface protein gene NcSRS2 and granule protein gene NcGRA7 were amplified by PCR,respectively.NcSRS2-NcGRA7 fusion gene was synthesized by SOE-PCR.The NcSRS2-NcGRA7 cloning vector was constructed and identified by PCR amplification,double digestion and bioinformatic analysis.The results showed that the sizes of NcSRS2,NcGRA7 and NcSRS2-NcGRA7 were 1061 bp,364 bp and 1482 bp,respectively.The cloning vector was demonstrated to be correct by PCR and double digestion.Sequencing analysis showed that the nucleotide sequences of NcSRS2 and NcGRA7 shared 99% homology with those published in GenBank(AF061249,AF176649).Prediction analysis by DNAman software indicated that the antigen index of NcSRS2-NcGRA7 was high.The main secondary structure of the fusion protein was mainly composed of α-helix and β-sheet.The tertiary structure analysis showed that the protein could fold into two independent regions connected with a linker.It indicated that the functions of NcSRS2 and NcGRA7 were not affected by each other.This study applies a foundation for the immunological functions of NcSRS2-NcGRA7 fusion protein of bovine N.caninum.

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