论文部分内容阅读
为了构建凤冈野生天麻的抗真菌肽基因(gaf)的表达载体,经BamHI和HindIII酶切的pMD18-T-gaf和pET32a(+)质粒的纯化回收产物并进行连接,连接产物pET32a(+)-gaf转化大肠杆菌感受态细胞DH5α后,采用菌落PCR、酶切鉴定和阳性质粒测序分析。结果表明:构建的凤冈野生天麻抗真菌肽基因表达载体通过菌落PCR、酶切鉴定和阳性质粒测序分析,目的片段成功插入载体,目的片段与预期的gaf基因大小(545 bp)相符。试验成功构建了贵州野生天麻抗真菌肽基因表达载体,pET32a(+)-gaf可以用来制备抗真菌蛋白。
In order to construct an expression vector for the antifungal peptide gene (gaf) of Phylloxia giganteum, the products of pMD18-T-gaf and pET32a (+) digested with BamHI and HindIII were recovered and ligated to ligate the product pET32a (+) -gaf transformed E. coli competent cells DH5α, colony PCR, enzyme digestion and positive plasmid sequencing analysis. The results showed that the Fungang wild Gastrodia antifungal peptide gene expression vector was successfully inserted into the vector by colony PCR, restriction enzyme digestion and positive plasmid sequencing. The target fragment was consistent with the expected gaf gene size (545 bp). Experiments successfully constructed Guizhou wild Gastrodia antifungal peptide gene expression vector, pET32a (+) - gaf can be used to prepare antifungal protein.