人新基因HBRP基因工程细胞系的建立

Construction of Genetically Engineered Cell Line of a Novel Gene HBRP in Human Testis

在研究牛精浆蛋白 (bovineseminalplasmaproteins ,BSPproteins)及其相关蛋白在受精卵发育过程中的重要作用时 ,在人睾丸组织中发现并克隆了一个与BSP蛋白相关新基因的序列 ,其开放阅读框架 (openreadingframe ,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型结构域 ,与BSP蛋白在结构上有一定的相似性 ,称其为人BSP相关蛋白 (humanBSP relatedprotein ,HBRP) ,并在GenBank注册 ,登录号为AF2 791 4。预测该蛋白是与BSP蛋白功能相关的结合蛋白 ,并已成功地将该基因定位在 1 9号染色体 1区 3带上。为进一步研究该新基因的生物学功能 ,利用重组质粒pEGFP C2 HBRP转染人胚肾HEK2 93细胞 ,通过G41 8筛选出药物抗性克隆 ,建立了稳定的基因工程细胞系 ,为该基因功能的研究奠定了基础 。

The novel gene HBRP which is related to bovine seminal plasma proteins(BSP proteins) in human testis was found and cloned when researching the important roles of BSP proteins in fertilization and the development of zygote.Its open reading frame(ORF) encoded a protein of 223 amino acids.There are 4 fibronectin Ⅱ-domains in the amino acid sequence,which is similar in structure to BSP proteins and called as HBRP(Human BSP-Related Protein,HBRP)and registered in GenBank,the landing number is AF27914.It is estimated that HBRP is a kind of binding protein which is similar in function to the BSP proteins.The new gene was successfully localized to human chromosome 19q13 by the technique of FISH.To further analyse the biological function of HBRP,stable cellar line a genetically engineered was constructed by the means of transfection of HEK293 cell by recombination pEGFP-C2/HBRP plasmid.This is very important to study the biological function of HBRP.;