关键词: 新生隐球菌, 酵母双杂交, cDNA文库

Abstract:

Objective To lay foundation for investigating interaction mechanism between Cryptococcus neoformans and host cells, yeast two-hybrid cDNA library of H99 cells was constructed. Methods mRNA was isolated from Cryptococcus neoformans (H99) in the mid-log phase. cDNA were then synthesized using biotin-conjugated Oligo (dT) primer in the 5’end. The double-strand cDNA was ligated to adapter and passed the cDNA size fractionation columns. The primary cDNA library was constructed by BP recombination and the cDNA expression library was constructed by LR recombination. Results The primary library contained a total clones of 1.13×10⁷ CFU with a recombination rate of 90.63%. The constructed cDNA library contained total clones of 1.25×10⁷ CFU, and the recombination rate was 100%. Conclusion A high-quality cDNA library of Cryptococcus neoformans was constructed successfully. These data might be useful for screening the host proteins interacting with Cryptococcus neoformans and for following functional studies of Cryptococcus neoformans in the future.

Key words: Cryptococcus neoformans, yeast two-hybrid, cDNA library