人Neuritin在原核表达系统的构建及表达纯化

Neuritin是一种新发现能促进神经突起和轴突分支的蛋白，为了更清楚的研究它的生物学功能，我们在已克隆Neuritin cDNA的基础上， PCR扩增出Neuritin ORF，与原核表达载体pET32a重组后，成功的构建了Neuritin原核表达质粒pET32a-Neuritin。重组质粒转化BL21大肠杆菌, IPTG诱导后,表达产物用SDS-PAGE 和Western blot证实系Neuritin，用镍离子亲和层析的方法获得了纯化的Neuritin蛋白。

Expressing Purification and Identification of Neuritin Gene in the E.coil

Neuritin is a new neurotrophic factor found recently. In order to identify the function of Neuritin clearly, the coding sequence of human neuritin was amplified by PCR from neuritin cDNA , this fragment digested by NocI and NotI was inserted into pET32a by T4 ligase and transformed into E. coli BL21 then the recombinant plasmid named pET32a-neuritin was constructed successfully . Neuritin was expressed distinctly after inducing by EPTG. The product was identified as neuritin by SDS-PAGE and Western blot analysis . The expression production was purified on Ni2 -NTA column.