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人骨形成蛋白2基因的克隆及序列测定
华 松1, 贾战生2, 武 浩1
1.西北农林科技大学 生物工程研究所;2.第四军医大学 唐都医院
摘要:
为了获得人骨形成蛋白2(hBMP-2)基因,以GeneBank(NM001200)中的hBMP-2基因编码序列为依据设计引物,采用RT PCR技术从人胎盘中获得hBMP-2基因cDNA,并将其重组到pMD18-T载体中,转化到大肠杆菌JM109后挑选阳性克隆,经双酶切鉴定并进行测序。结果表明,获得的基因片断为hBMP-2全编码序列,同源性为99.9%。
关键词:  人骨形成蛋白2  RT-PCR  克隆
DOI:
分类号:
基金项目:国家“863”高技术资助项目(2001AA213081)
Cloning and sequencing of Human Bone Morphogenetic Protein 2
Abstract:
In order to clone the human bone morphogenetic protein 2 (hBMP-2),a pair of primers were designed based on the coding sequence of hBMP-2 derived from the GeneBanks (NM001200).The total RNA was extracted from the tissue of human placenta.The target fragment was amplified by RT-PCR,then inserted into cloning vector pMD18-T and transformed into E.coli.strain JM109.The result of double restriction endonuclease digestion and the sequencing indicated that the fragment cloned was correct compared with the sequence deposited in GeneBanks,and the homology was 99.9%.Cloning of hBMP-2 gene gives a basis for furher expression study.
Key words:  hBMP-2,RT-PCR,cloning