人K562细胞cDNA文库的扩增、纯化、鉴定和酵母细胞转化

Amplification,purification and identification of human K562 cell cDNA library and its transformation into yeast cell

目的:对预转化到大肠杆菌DH5α中的人K562细胞cDNA文库进行扩增、纯化和鉴定并将文库质粒转化酵母细胞,为下一步的靶蛋白筛选做准备。方法:对K562细胞cDNA文库进行扩增,提取文库质粒,将文库质粒转化酵母Y187细胞,并用PCR和酶切鉴定转化结果。结果:成功的扩增人K562细胞cDNA文库、并验证了文库的多样性、将文库质粒成功转化酵母Y187细胞。结论:K562细胞cDNA文库的成功扩增、纯化、鉴定,为进一步的文库筛选奠定了基础。

Objective：To Amplify,purify and identify the cDNA library of human K562 cell in E.coli DH5α and to transform it into yeast cell for subsequent target protein screening. Methods：The cDNA library of human K562 cell was amplified. Then the library plasmids were extracted and transformed into Y187 yeast cell. The results of transformation were verified by PCR and restriction enzyme analysis. Results：The cDNA library of human K562 cell was amplified successfully and the diversities were verified. The cDNA library of human K562 cell was also successfully transformed into Y187 yeast cell. Conclusion： Successfully amplification,purification and identification of the eDNA library of human K562 cell lay the foundation for the subsequent protein screening.