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人牛精浆蛋白相关新基因的cDNA克隆、定位和表达
引用本文:罗阳,张学,刘莹,姜莉,刘兴元,于秉治.人牛精浆蛋白相关新基因的cDNA克隆、定位和表达[J].中国生物化学与分子生物学报,2002,18(2):191-196.
作者姓名:罗阳  张学  刘莹  姜莉  刘兴元  于秉治
作者单位:1. 中国医科大学基础医学院生物化学教研室,沈阳,110001
2. 中国医科大学卫生部细胞生物学重点实验室,沈阳,110001
基金项目:973国家基础研究重点课题资助 (No .19990 5 5 90 0 2 ),国家自然科学基金重点课题资助 (No .39730 46 0 )~~
摘    要:为了研究牛精浆 (bovineseminalplasma ,BSP)蛋白及其相关蛋白在受精及受精卵发育中的重要作用 ,寻找BSP蛋白相关新基因 .采用cDNA末端快速扩增 (RACE)技术 ,克隆了一个BSP蛋白相关基因的cDNA序列 .应用辐射杂种细胞系 (RH)技术进行了基因染色体定位 .通过RT PCR检测了该基因在人体各组织中的表达情况 .并将该基因编码的蛋白进行了原核表达 .新基因的cDNA长度为 10 5 2bp ,其开放阅读框架 (ORF)编码了一个含 2 2 3个氨基酸残基的蛋白质 ,氨基酸序列中含有 4个纤连蛋白Ⅱ结构域 ,与BSP蛋白在结构上具有一定的相似性 ,称其为人BSP相关蛋白 (humanBSP relatedproteins ,HBRP) .该基因定位于染色体 19q13,在大肠杆菌中表达为 5 2kD的融合蛋白 .研究结果提示 ,应用RACE方法克隆了一种新的人类与BSP蛋白相关的基因 ,推测其编码蛋白是与BSP蛋白功能相关的结合蛋白 ,通过基因重组技术大量获得表达蛋白 ,对进一步研究新蛋白的生物学功能具有重要的意义 .

关 键 词:牛精浆  人牛精浆相关蛋白  cDNA末端快速扩增  基因克隆与定位  GST融合表达  
收稿时间:2002-04-20
修稿时间:2001年9月27日

Cloning,Location and Expression of cDNA for a Noval Gene Related to BSP Protein in Human
LUO Yang ,ZHANG Xue ,LIU Ying ,JIANG Li ,LIU Xin-yuan ,YU Bing-zhi.Cloning,Location and Expression of cDNA for a Noval Gene Related to BSP Protein in Human[J].Chinese Journal of Biochemistry and Molecular Biology,2002,18(2):191-196.
Authors:LUO Yang  ZHANG Xue  LIU Ying  JIANG Li  LIU Xin-yuan  YU Bing-zhi
Institution:( 1) Department of Biochemistry, 2) Department of Cell Biology,China Medical University, Shenyang 110001,China
Abstract:To find a novel gene which is related to protein of bovine seminal plasma (BSP) in human and study the new gene′s structural feature,expression and location in chromosome,the cDNA was cloned by using the 3′ rapid amplification of cDNA ends (RACE) and 5′ RACE methods and cloned into the expression vector of GST fusion protein (PGEX-5X-1),finally expressed in E.coli BL 21. The location of the new gene in the human chromosome was determined by the technique of radiation hybrid(RH).The cDNA was 1 052 bp long and its open reading frame (ORF) encoding a protein of 223 amino acid.Further analyisis showed that there were 4 fibronectin type Ⅱ-medules in the amino acid sequence,which was similar to BSP protein as far as structure was concerned.The expression product of the plasmid which was constructed of GST fusion protein was a fusion protein of approximate 52 kD.The new gene was successfully located on human chromosome 19q13 by the technique of RH.The results suggested that a new gene HBRP (human BSP-related protein) was cloned.The function of HBRP is related to that of BSP protein family and the HBRP gene is located in 19q13.Moreover,a great amount of HBRP protein could be obtained by the technique of gene recombination,it provided a reliable foundation to investigate the biological function of this new protein.
Keywords:bovine seminal plasma(BSP)  humam BSP-related protein    RACE  gene cloning and location  GST fusion expression
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