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《The Journal of Bone and Joint Injury》 2003-09
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Molecular Cloning and Sequence Analysis of Full - Length cDNA Encoding Human Bone Morphogenetic Protein - 7

Li Xinyou , Liu Miao , Wang Yi, et al. Department of Orthopedics of First Hospital of Xi'an JiaoTong University , 710061  
Objective To clone the full-length human bone morphogenetic protein-7 (BMP- 7) gene and analyses its sequence, to aid in investigation of its function and structure. Methods Total RNA was isolated from Chinese fetal kidney by the acid guanidinium thio-cyanate phenol-chloroform method. Two overlapping segments of human BMP - 7 cDNA were obtained by reverse transcription (RT) - PCR. Following application, the two segments were ligated to each other and subcloned into PGEM - T easy vector to form PEGM -T easy/hBMP-7 recombinant plasmid. Sanger dideoxy chain-termination method Sanger, 1977 # 423 was used to sequence the cD-NA. Results 750 bp fragment from 5' end of BMP- 7 gene was prepared by RT- PCR using # 2 primer (PCR by using #2 and # 1), and 540bp fragment from 3'end was generated by RT - PCR using # 4 primer ( PCR using # 3 and # 4). Full-length cDNA encod-, ing BMP- 7 was obtained by religation of two segments. When compared with hBMP - 7 sequence in Genebank (XM30619) , our full-length BMP - 7 cDNA has a G instead of a T at nucleotide 842. This change results in valine substituting for phenylalanine in the protein. Conclusion This is the first time that BMP - 7 cDNA was successfully cloned from Chinese fetal kidney. BMP - 7 plays an important role in healing injuries of the osteo-articular system. This makes BMP - 7 is an attractive target for various clinical applications.
【CateGory Index】: R346
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