树鼩HGF全长编码序列的克隆及分子特征分析

Cloning of full-length coding sequence of tree shrew HGF and prediction of its molecular characteristics

目的获取树鼩肝细胞生长因子的全长编码序列并进行分子特征分析。方法以树鼩肝组织总RNA为材料,通过RT-PCR扩增和序列拼接获得树鼩肝细胞生长因子全长编码序列,进而通过DNAMAN,Pymol等生物信息学软件对其序列和分子特征进行分析。结果树鼩HGF分子结构在进化中相对保守,树鼩HGF整体结构与人相似。但树鼩HGFαNK1区域相比人HGF多出一个α螺旋,并且有两个更长的β折叠片,另外,树鼩在该区域存在一个很强的表面正电荷区域。这些差异可能会对树鼩HGF与其受体的作用方式产生影响。结论本实验为明确树鼩HGF与其受体的作用方式和树鼩HGF的合成与制备奠定了基础。

Objective To get the full-length HGF,Hepatocyte growth factor, cDNA encoding sequence and predict its molecular characteristics. Methods Full-length HGF cDNA encoding sequence is cloned from total RNA in liver tissue of tree shrew by RT-PCR and its molecular characteristics is analyzed compared with other mammals by using biology software such as DNAMAN,Pymol and so forth. Results A 2193bp of the full-length HGF cDNA encoding sequence is cloned from total RNA in liver tissue of tree shrew. The analysis of molecular characteristics show that the molecular structure of HGF is relatively conservative in evolution, tree shrew HGF overall structure similar to Homo sapiens. However,compared with the structure and charge surface of NK1 domain of HGFα-chain of human ,tree shrew has two longer β-sheet and one more α-helix ,Additional,tree shrew had a strong positive charged site in this area,which may have a impact on the bind of HGF to its receptor. Conclusion This study provide a theoretical basis for preparation and receptor-binding mechanism studies of tree shrew HGF.