中文摘要：

目的构建含小鼠MASH1基因的绿色荧光蛋白表达载体pEGFP-C3-MASH1，为进一步研究MASH1在间充质干细胞神经分化中的作用打下基础。方法应用RT-PCR方法从小鼠13．5d胚胎组织中扩增出两端带有HindⅢ和EcoRI酶切位点的MASH1cDNA编码片段，经回收、纯化、酶切后，依次连接到质粒pGEM-T和pEGFP-C3上。结果酶切及测序结果表明：重组质粒PEGFP-C3含有MASH-Ⅰ片段，方向及大小正确。结论成功构建了小鼠真核表达载体PEGFP-C3-MASH-Ⅰ。

英文摘要：

Objective To construct eukaryotic fluorescent expression vector pEGFP - C3 - MASH1 for studing the effect of MASH1 on the neural differentiation of stem cells. Methods The cDNA coding sequence of MASH1 containing the digestion site of Hind Ⅲ and EcoRI was amplified by RT - PCR from the mouse embryo and then was inserted into pGEM - T plasmid and pEGFP - C3 fluorescent expression vector . The recombined plasmid was identified by restriction endonuclease digestion ,PCR technique and sequencing. Results Restriction erolonuclease digestion and DNA sequencing results showed that the recombinant plasmid PEGFP-C3 contained the fragment of MASH-Ⅰ. Conclusion The enkaryotic expression vector PEGFP-C3-MASH-Ⅰ was constructed sucessgully.