Cloning of integral mature peptide gene of human GDF-5.
The integral mature peptide gene of human growth differentiation factor-5 (GDF-5) was cloned to provide the essential foundation for study on the biological characteristics of GDF-5 at gene and protein levels. Two primers were chemosynthesized according to the hGDF-5 sequence reported in Genbank. The hGDF-5 gene was gained by RT-PCR methods from the total RNA extracted from human fetus cartilage tissue, and was cloned into vector pMD18-T. The sequence of recombinant plasmid pMD18-T-hGDF-5 was analyzed by sequence analysis. DNA agarose gel electrophoresis showed that the product of RT-PCR was about 380bp, and double enzyme digestion of the recombinant plasmid corresponded with it. The result of sequence assay was in agreement with the reported hGDF-5 sequence in Genbank. Our results showed that the integral mature peptide gene of human GDF-5 was cloned successfully from human fetal cartilage tissue, and totally identified with the sequence of human GDF-5 in Genbank.
Wang
Wanshan
W
Department of Anatomy, First Military Medical University, Guangzhou 510515, China.
Gu
Weiwang
W
Wang
Qiwei
Q
Piao
Zhongxian
Z
Piao
Yingjie
Y
D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000235:genetics; D002356:Cartilage / Q000737:chemistry; D003001:Cloning, Molecular; D005333:Fetus; D005822:Genetic Vectors; D055428:Growth Differentiation Factor 5; D006801:Humans; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000235:genetics
D019485:Bone Morphogenetic Proteins; C525893:GDF5 protein, human; D055428:Growth Differentiation Factor 5; D016212:Transforming Growth Factor beta