人骨形成蛋白3基因cDNA部分序列的RT-PCR克隆  

Cloning of partial cDNA of human bone morphogenetic protein 3 (hBMP-3) gene by RT-PCR

作　　者：张涌[1] 戴博[1] 刘建忠[1] 郑月茂[1] 

机构地区：[1]西北农林科技大学生物工程研究所,陕西杨陵712100

出　　处：《西北农林科技大学学报（自然科学版）》2003年第2期1-5,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家 8 63高技术计划项目 (2 0 0 1AA2 13 0 81)

摘　　要：针对人骨形成蛋白 3基因的 c DNA序列设计引物 ,通过反转录 PCR技术 ,从人骨纤维肉瘤组织中扩增出目的片段。从胶中回收目的片段 ,将其与 p MD18- T载体连接 ,转化大肠杆菌 DH 5α后 ,经 H ind 酶切鉴定 ,进行测序。结果显示所得到的片段大小为 84 5 bp,与所设计的序列同源性为 10 0 % ,说明已经成功地扩增、克隆了人骨形成蛋白 3(h BMP- 3)基因 c DNA的大部分序列。A pair of primers for the amplification of partial cDNA sequence of human bone morphogenetic protein 3 gene were designed,the target fragment was amplified by RT PCR from the tissue of human bone fiber sarcoma,then was purified from agarose gels and integrated into pMD18 T vector.After being transformed into E.coli. DH5α,tested by Hin dⅢ restriction endonuclease digestion,the positive clone was sequenced.The sequencing result indicated that the size of the fragment cloned is 845 bp,and its sequence homology with the designed one is 100%.The conclusion can be made that the major part of the cDNA of human bone morphogenetic protein 3 (hBMP 3) gene has been successfully amplified and cloned.

关 键 词：人骨形成蛋白3基因 CDNA序列 RT-PCR 基因克隆 骨形成蛋白 转基因动物乳腺 

分 类 号：Q785[生物学—分子生物学] R394.8[医药卫生—医学遗传学]