Hu Shenglin, Wang Zhefan, Wang Dongmei, Wang Jichao, Hong Jiong
School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027 People's Republic of China.
Hefei National Laboratory for Physical Science At the Microscale, Hefei, Anhui 230026 People's Republic of China.
3 Biotech. 2021 Sep;11(9):414. doi: 10.1007/s13205-021-02963-w. Epub 2021 Aug 18.
is a thermophilic fungus that belongs to the ascomycetous class and has attracted increasing interest for its ability to produce thermostable cellulolytic enzymes and growth at elevated temperatures. However, studies on this organism have been limited because of the lack of a genetic manipulation system. Here, we developed a polyethylene glycol (PEG)-mediated transformation system for based on an orotidine-5'-monophosphate decarboxylase (pyrG)-deficient mutant, with this method achieving a transformation efficiency of 33 ± 3 transformants per microgram of DNA. Intracellular or secretory expression of heterologous proteins, including green fluorescent protein, -galactosidase and -amylase, in was successful under the inducible endogenous cellobiohydrolase and endoglucanase gene promoter or the constitutive heterologous pyruvate decarboxylase and enolase gene promoter from . To the best of our knowledge, this is the first report on PEG-mediated transformation of , which sets the foundation for strain improvement for biotechnological applications and functional genomic studies.
The online version contains supplementary material available at 10.1007/s13205-021-02963-w.
是一种嗜热真菌,属于子囊菌纲,因其产生热稳定纤维素分解酶的能力和在高温下生长的特性而越来越受到关注。然而,由于缺乏遗传操作系统,对这种生物体的研究一直有限。在此,我们基于尿苷-5'-单磷酸脱羧酶(pyrG)缺陷型突变体开发了一种用于的聚乙二醇(PEG)介导的转化系统,该方法实现了每微克DNA 33±3个转化子的转化效率。在来自的诱导型内源性纤维二糖水解酶和内切葡聚糖酶基因启动子或组成型异源丙酮酸脱羧酶和烯醇酶基因启动子的作用下,在中成功实现了包括绿色荧光蛋白、β-半乳糖苷酶和α-淀粉酶在内的异源蛋白的细胞内或分泌表达。据我们所知,这是关于PEG介导的转化的首次报道,为生物技术应用和功能基因组学研究的菌株改良奠定了基础。
在线版本包含可在10.1007/s13205-021-02963-w获取的补充材料。