三孢布拉霉中光受体的克隆表达和生物信息学分析

Light can induce accumulation of carotenoids in the mycelium of Blakesleatrispora and affect other phenotypes, but the mechanism of light sensing in B. trispora are not clear so far. In this study, based on the conserved sequences of photoreceptors in many closely related filamentous fungi, the full-length sequences and coding sequences of two genes of btwc-1a and btwc-1c were obtained by chromosome walking and RT-PCR technologies. The bioinformatics analysis showed that BTWC-1A had the highest identity (87%) with the WC-1 of Choanephoracucurbitarum, while BTWC-1C had the highest identity (65%) with the WC-1C of Mucorcircinelloides. The functional domains of BTWC-1A and BTWC -1C contain 2-3 PAS structures and a zinc finger structure, and the N-terminal LOV domain is highly conserved as those in the photoreceptors of many other filamentous fungi. At the same time, the prokaryotic expression vectors pET28a-wc-1a and pET28a-wc-1c were constructed and the recombinant bacteria were induced by IPTG. Electrophoretic analysis showed that the molecular weights of the expressed proteins were about 35 kDa. This study was to clarify the process and molecular mechanism of light sensing in B. trispora and lay a foundation for the regulation of carotenoid synthesis by light.