南京医科大学学报（英文版）2003，Vol.17Issue(2)：62-66,5.
人骨形态发生蛋白-7全长基因cDNA的克隆和序列分析
Molecular Cloning and Sequence Analysis of Full-Length cDNA Encoding Human Bone Morphogenetic Protein-7
李新友 1刘淼 1李曙明 1姚煜 1王全颖 2杨广笑2
作者信息
- 1. 西安交通大学第一医院骨科,西安,710061
- 2. 西安华广生物工程公司,西安,710075,中国
- 折叠
摘要
Abstract
Objective: To clone the full-length human bone morphogenetic protein-7 ( BMP-7) gene and analyse its sequence, to aid in investigation of its function and structure. Methods: TotalRNA was isolated from Chinese fetal kidney by the acid guanidinium thiocyanate phenol-chloroformmethod. Two overlapping segments of human BMP-7 cDNA were obtained by reverse transcription(RT)-PCR. Fallowing application, the two segments were ligated to each other and subcloned intoPGEM-7 easy vector to form PEGM-T easy/hBMP-7 recombinant plasmid. Sanger dideoxy chain-ter-mination method was used to sequence the cDNA. Results: There was 750 bp fragment obtained RT-PCR using # 2 primer from 5' end of BMP-7 gene (PCR by using # 2 and # 1) ,and 540 bp frag-merit from 3' end was generated by RT-PCR using # 4 primer (PCR using # 3 and # 4). Full-lengthcDNA encoding BMP-7 was obtained by religation of two segments. When compared with hBMP-7 se-quence in Gene bank (XM30619) ,our full-length BMP- 7 cDNA has a G instead of a T at nucleotide862. This change results in valine substituting for phenylalanine in the protein. Conclusion: This isthe first time that BMP-7 cDNA was successfully cloned from Chinese fetal kidney. BMP-7 cDNAplays an important role in healing injuries of the osteo-articular system. This makes BMP-7 is an at-tractive target for various clinical applications.关键词
骨形态发生蛋白-7/基因克隆/序列分析Key words
bone morphogenetic protein-7/gene clone/sequence分类
医药卫生引用本文复制引用
李新友,刘淼,李曙明,姚煜,王全颖,杨广笑..人骨形态发生蛋白-7全长基因cDNA的克隆和序列分析[J].南京医科大学学报（英文版）,2003,17(2):62-66,5.
