黄鳝生长激素基因cDNA的克隆及胞内真核表达载体的构建

为大量获得具有生物学活性的黄鳝生长激素，采用RT-PCR的方法从黄鳝脑cDNA中扩增得到了黄鳝生长激素基因。通过特异性的引物扩增后，将黄鳝生长激素基因ORF区域序列连接到酵母胞内表达载体pGAPZB中，构建了重组胞内表达载体pGAPZB—rfeGH。用BlnI线性化重组质粒后，电击转化毕赤酵母GS115。经过Zeocin筛选和PCR扩增转化子基因组筛选，得到了数株基因工程酵母菌，序列测定表明生长素基因已经成功的转移到酵母基因上。黄鳝生长激素胞内表达载体的构建成功为进一步研究黄鳝生长激素基因的功能和大量开发并利用奠定了实验基础。

Molecular Cloning and Construction of Expression Vector of Growth Hormone Gene of Rice Field Eel (Monopterus albus)

Growth hormone plays an essential role in the stimulation of somatic growth and development. To produce large amount of rice field eel growth hormone （rfe-GH） of biological activity for further use, this study aimed at high yield expression of rfe-GH in the yeast Pichia pastoris. The rfe-GH cDNA was amplified by RT-PCR and inserted into a yeast vector pGAPZB. Subsequently, the vector was digested with BlnI and transformed into GS115. After screening on plates with Zeocin, PCR was performed to amplify and identify the genomic DNA of these transformants. Finally, sequencing of the transformants suggested that the rfe-GH had been transferred into the genome of the yeast. The construction of the vector provide us basic data for further study of function of the rfe-GH.