title: [Cloning, expression analysis and RNA interference of FGF5 gene in sheep].
abstract: The cDNA of fibroblast growth factor 5 (FGF5) gene in sheep was cloned, and the nucleotides sequence homology of FGF5 was compared with other six mammal. In addition, the expression of FGF5 in different tissues was analysed. Gene FGF5 was then recombined into prokaryotic expression vector (pGEX-4T-2) and RNA interference vector (pSilencer 5.1 H1) to study its expression in fibroblast cell lines. Results showed that the open reading frame (ORF) of cDNA in sheep consisted of 813 nucleotide acids encoding 270 amino acids, with the molecular mass of 29.58 kDa and theoretical pI of 10.59. The amino acids sequence of FGF5 gene in sheep shared high identity with those in cow, human, mouse, rat, dog, cat and rabbit. In addition, analysis on tissue expression showed that FGF5 expressed in skin, heart, kidney, liver, pancreas, spleen, lung, and small intestine, especially presenting high levels in skin. The expression of FGF5 in E. coli was induced with IPTG, which produced a protein band with the expected size of 56 kDa on SDS-PAGE, while the expression of FGF5 in sheep fibroblast cell line was knocked down remarkably with the help of integrated RNAi vector.
authors.affiliation: College of Animal Science, Shihezi University, Shihezi, China. lwx_666@sina.com
authors.forename: Wu-Xian
authors.initials: WX
authors.lastname: Liu
authors.forename: Bin
authors.initials: B
authors.lastname: Jia
authors.forename: Guo-Qing
authors.initials: GQ
authors.lastname: Shi
authors.forename: Jian-Gong
authors.initials: JG
authors.lastname: Ren
authors.forename: Ka
authors.initials: K
authors.lastname: Liu
authors.forename: Run-Lin
authors.initials: RL
authors.lastname: Ma
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002415:Cats; D002417:Cattle; D003001:Cloning, Molecular; D004285:Dogs; D051521:Fibroblast Growth Factor 5 / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010802:Phylogeny; D034622:RNA Interference; D051381:Rats; D016415:Sequence Alignment; D012756:Sheep
chemical_list: D051521:Fibroblast Growth Factor 5