Application of conditional promoter pCTR4 in Cryptococcal gene expression regulation

Abstract

Abstract: Objective To explore the application of conditional promoter pCTR4 in Cryptococcal gene expression regulation via construction of new vector pNEO/CTR4 and promoter replacement.Methods Constructed the new plasmid pNEO/CTR4 fusing report gene NEO and copper-repressive promoter pCTR4 via overlap PCR.On the basis, constructed its promoter replacement cassette, and then transformed them into Cryptococcal competent cells via gene gun to get its reconstitution strains, which were tested via regular and real-time PCR.Results Correct new vector pNEO/CTR4 and Cryptococcal reconstitution strains were obtained and confirmed.Conditional promoter pCTR4 played induced or repressed effect in UBI1 transcription under different media as expected.Conclusion The new vector of copper-repressed promoter was applicable in regulating Cryptococcal gene expression.And the ubiquitin encoding gene UBI1 was not essential for viability in Cryptococcus neoformans.Our study laid the groundwork for following functional studies of ubiquitin system in Cryptococcus neoformans.

Key words: Cryptococcus neoformans, conditional promoter, ubiquitin, pathogenesis

CLC Number:

R379.5

ZHAO Jing-yu, FA Zhen-zong, FANG Wei, MENG Yun-fang, ZHANG Chao, ZHOU Zhao-jing, PAN Wei-hua, LIAO Wan-qing. Application of conditional promoter pCTR4 in Cryptococcal gene expression regulation[J]. Chinese Journal of Mycology, 2014, 9(1): 1-6.

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