大鼠神经突起因子的克隆及表达载体的构建

Construction of Cloning and Expression Vector of Rat Neurtin cDNA

从成年SD大鼠脑组织分离提取总RNA,反转录PCR扩增出鼠神经突起因子(neuritin)cDNA的最大开放阅读框(openingreadingframe,ORF)后,重组于克隆载体pGEM3z上,经PCR、限制内切酶鉴定和DNA测序鉴定,扩增出的cDNA426bp全部核苷酸序列与国外文献报道的完全一致;在已构建好的重组克隆载体neuritin pGEM3z基础上,将该neuritinORF亚克隆到原核表达载体pQE30上,为获取具有天然活性的neuritin蛋白及其功能研究提供了基础。

Total RNA were isolated from the brain of an adult SD rat. The opening reading frame (ORF) of neuritin cDNA was amplified by the reverse transcription polymerase chain reaction (RT-PCR). The cDNA was inserted into clonig vector(pGEM3z), and identified by PCR restriction endonuclease and sequencing. The result of DNA sequencing was successful.On the basis of recombinant neuritin-pGEM3z vector, we subcloned the neuritin cDNA into pQE30 which is a prokaryotic expression vector in order to obtain the natural Neuritin protein, and then do the functional research of it.