产紫杉醇真菌NK-101中PAM基因的功能性研究
he Functional Research on PAM Gene of Taxol-Producing Fungus NK-101
本实验室保藏有一株具有产紫杉醇能力的拟盘多毛孢菌NK-101，目前已完成全基因组测序。在此基础上，本实验室通过氨基酸序列比对，得到了NK-101菌株中与加拿大红豆杉（Taxus canadensis）中苯丙氨酸氨基变位酶（phenylalanine aminomutase）（GenBank：AAT47186.1）同源性较高的基因，将其命名为PAM基因；通过农杆菌介导的丝状真菌转化的方法，将敲除质粒转入NK-101菌株中并将PAM基因定向敲除，获得△pam菌株。通过对比野生型NK-101菌株和敲除菌株的表型，发现PAM基因的缺失导致敲除菌株生长速率变慢、菌体生物量减少、产孢能力下降、色素产生能力降低，菌株的次级代谢水平发生显著性下调，多种次级代谢产物基本检测不到。以上结果表明，PAM基因对菌株的生长、产孢及次级代谢水平具有一定的负调节作用。
We have isolated a taxol-producing fungus -Pestalotiopsis malicola named NK-101, and finished the whole genome sequencing and annotating work of NK-101. Comparing with the amino acid sequences of phenylalanine aminomutase (GenBank: AAT47186.1) in Taxus canadensis, we have found the homologous gene in NK-101 and named it PAM gene. Based on the homologous recombination and Agrobacterium-mediated transformation, we constructed pam gene deletion strain, named △pam. Comparing to NK-101, the growth rate, asexual spore production capacity, pigment secretion capacity, mycelial biomass and secondary metabolites synthesis capacity have decreased in △pam. Based on the above factors mentioned, we demonstrated that the PAM gene could negatively regulate the growth, asexual spore production, pigment secretion and secondary metabolites in NK-101.
马天竺、王宇、潘皎、朱旭东、李强、颜菲
生物科学研究方法、生物科学研究技术微生物学分子生物学
拟盘多毛孢菌PAM基因丝状真菌转化基因敲除
Pestalotiopsis malicolaPAM geneAgrobacterium-mediated transformationgene knockout
马天竺,王宇,潘皎,朱旭东,李强,颜菲.产紫杉醇真菌NK-101中PAM基因的功能性研究[EB/OL].(2013-07-17)[2025-08-16].http://www.paper.edu.cn/releasepaper/content/201307-253.点此复制
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