摘要
目的克隆人骨形态发生蛋白-7成熟肽基因。方法根据Genebank人骨形态发生蛋白-7基因序列合成两条引物,从培养的人胎儿软骨细胞中提取mRNA,利用OnestepRT-PCR技术扩增出人骨形态发生蛋白-7成熟肽的基因序列,将PCR扩增产物基因片段连接克隆载体质粒中转化大肠杆菌DH5α进行克隆筛选鉴定及测序。结果琼脂糖凝胶电泳检测RT-PCR产物显示一长约451bp的条带,阳性克隆质粒经双酶切可切出约451bp的片段,DNA测序结果与Genebank中的序列相符。结论利用RT-PCR技术可成功的从人胎儿软骨细胞中克隆出人BMP-7成熟肽基因。
[Objective] To clone the eDNA of mature peptide of human bone morphogenetic protein-7. [Methotis] According to the hBMP-7 sequence reported in Genebank , two primers were designed. The mature peptide of hBMP-7 eDNA was gained by onestep RT-PCR from the mRNA, Which was extracted from culture cartilage ceils of rectus. Then it was cloned into the vector of PGEM-T and the result plasmid was transformed into DH5cα. The sequence of the plasmid BMP-7 was detected by DNA sequence machine. [Results] DNA agarose electrophoresis showed that the product of RT-PCR was about 450bp, it was corresponed with the result of two enzymed digestion of the recombinanted plasmid. The result of sequence was identical with the reported hBMP-7 sequence in Genebank.[Condusion] The mature peptide of hBMP-7 eDNA can be cloned successfully from cartilage ceils of foetus.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2005年第14期2143-2144,2149,共3页
China Journal of Modern Medicine
关键词
骨形态发生蛋白
克隆
基因
bone morphogenetic protein
cloning
gene
