Abstract:Abstract: In order to study the expression and regulation of cellulase gene in Trichoderma reesei, overlap PCR amplification and molecular cloning techniques were employed to construct a vector pLXT-ZsGreen expressing bright green fluorescence protein in T. reesei. The vector contains prokaryotic replication origin ColE1, the ampicillin resistance gene, promoter of pyruvate decarboxylase (PDC), terminator of PDC, ZsGreen gene, and the hygromycin B resistance gene. The vectors were transformed into the protoplasts of T. reesei QM9414. The hyphae of the transformants were observed using a fluorescence microscope with 488 nm excitation light, and four colonies were randomly selected for Western-blot. The results showed that the transformants were able to express bright green fluorescence under fluorescence microscope. The Western-blot analysis further verified that the constructed plasmid could express ZsGreen protein effectively in T. reesei. The vector pLXT-ZsGreen is capable of stable and efficient expression of exogenous genes in T. reesei, and this lays a foundation for the study of gene expression regulation of T. reesei.
引用本文:
高云雨, 佘炜怡, 董冠园, 钟路遥, 刘雯莉, 田生礼. 里氏木霉绿色荧光蛋白表达载体的构建及功能鉴定[J]. 生命科学研究, 2017, 21(4): 306-311.
GAO Yun-yu, SHE Wei-yi, DONG Guan-yuan, ZHONG Lu-yao, LIU Wen-li, TIAN Sheng-li. Construction of ZsGreen Expression Vector and Verification of Its Function in Trichoderma reesei. Life Science Research, 2017, 21(4): 306-311.