一种新发现神经突起生长诱向因子的全长克隆及功能研究

Hetrin gene was a new member of the netrin family with axon guidance function. In order to study the function of hetrin in nervous system, the full-length cDNA sequence of hetrin was obtained at first. After then, the gene structure, chromosomal localization, alternative splicing form of the hetrin gene was studied systematically. Based on multiple tissues Dot blot and Northern blot technique, the tissue expression profile of the hetrin gene was carefully studied. Furthermore, we used the in situ hybridization technique to study the exact distribution of the hetrin mRNA in the rat brain. At the same time, the hetrin protein with biological activity was obtained using bioengineer technique using both prokaryotic expression vector (pGEX-4T-2) and eukaryotic expression vector(pcDNA3.1). Moreover, we found that hetrin could bind and interact with another important nervous system specific gene-neuronatin using yeast two hybridization technique. This has shed much more lights on the functional study of the hetrin protein. It is believed that further study on the hetrin gene and its interacting gene will break through the understand of the mechanism of development, axon guidance and plasticity of the nervous system. We wish this project to be continued under further support of the NSFC.

围绕我们新近发现并率先向GenBank数据库注册的肝源性神经突起生长诱向因子（Hetrin）醒芯浚獠捎肦ACE技术得到其全长cDNA，进而构建其表达载体并测活，采用Northern Blot、原位杂交等技术研究Hetrin基因在大鼠神经系统发育过程中的表达图谱，进而探讨其与大鼠神经系统再生的关系，研究该因子对神经突起生长的促进作用，从而为系统地认识其功能奠定基础。