山羊IGFBP-5基因克隆及特征分析

为探讨山羊IGFBP-5基因的特点,进一步研究其在山羊上的表达及生长发育中的作用机制,以初生南江黄羊肝脏组织为材料抽提RNA进行克隆测序,并利用ExPasy、NetPhos、NetOGlyc、SignalP等生物软件分析IGFBP-5基因CDS区及其编码氨基酸的理化性质、结构特点.结果显示,山羊IGFBP-5基因的CD...

Molecular Cloning and Characterization of Goat IGFBP-5 Gene

In order to reveal the construction features of goat IGFBP-5 and provide a basis for further study on its expression and physical function in goat growth and development,IGFBP-5 CDS was cloned from liver of newborn Nanjiang Mongolian Grazelle,and analyzed using biology softwares,for instance:ExPasy,NetPhos,NetOGlyc,and SignalP.The whole length of IGFBP-5 CDS was 819bp(GC 64.96%) and encoded 272 amino acid residues.The theory values of IGFBP-5 pI and molecular weight were 8.56 and 30.392kD,respectively.Residues 1 to 19 at N-terminal formed signal peptide,and there were two hydrophilic regions and five hydrophobic regions in primary sequence of goat IGFBP-5,while the grand average of hydroppathicity(GRAVY) was-0.619,and sub-cellular localization was extracellular for mature peptide.Followed by alpha helix(13.6%) and extended strand(2.6%),random coil(83.8%) was most common.Insulin-like growth factor-binding protein(IGFBP) N-terminal domain signature and thyroglobulin type-1 repeat signature in C-terminal region were found.There were 3 O-glycosylation sites and 22 phosphorylation sites,and the main modification included CKⅡ,PKA,PKC,and MAPK.The homology of nucleotide and amino acid sequence of goat IGFBP-5 gene with others was above 89% and 94%,respectively,in addition,the N-terminal sequences were same among species.There was one particular amino acid insertion in primary IGFBP-5 of human(Leu3) and goat(Ala102).The results indicated that IGFBP-5 of goat was a weakly hydrophilic secretory protein with signal peptide,and phosphorylation was the main factor to regulate its function.