粉尘螨Der f15的基因克隆与其表达载体的构建
Cloning and Vectorconstuction of Der f15 from House Dust Mite Dermatophagoides Farina

先挑取纯培养的粉尘螨,提取总的RNA,后采用RT-PCR方法进行反转录出cDNA.由cDNA提取出目的基因进行片段扩增,产物连接入T载体(pMD18-T).经扩增后,用EcoR I和Xho I双酶切,将目的基因分别连接到pET28和pET32的表达载体上,得到重组质粒pET28和pET32,即粉尘螨的基因克隆与 表达载体构建成功.
The living mites of south China area which had been identified and cultured were picked,from which extracted the total RNA.RT-PCR technology were used to clone the gene and then connected it to the T vector(pMD18-T).After that,the expression vector(pET28 and pET32)was constructed for the new gene segment and were connect ed together