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产紫杉醇真菌NK-101中PAM基因的功能研究

发表时间:2014-03-15  浏览量:1387  下载量:561
全部作者: 颜菲,王宇,李强,马天竺,朱旭东,潘皎
作者单位: 南开大学生命科学学院
摘 要: 本实验室保藏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,通过氨基酸序列比对,得到了NK-101菌株与加拿大红豆杉(Taxus canadensis)中苯丙氨酸氨基变位酶(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Δpam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基因与NK-101菌株的生长、产孢及次级代谢水平密切相关。
关 键 词: 分子生物学;拟盘多毛孢菌;PAM基因;丝状真菌转化;基因敲除
Title: Functional research on PAM gene of taxol-producing fungus NK-101
Author: YAN Fei, WANG Yu, LI Qiang, MA Tianzhu, ZHU Xudong, PAN Jiao
Organization: College of Life Sciences, Nankai University
Abstract: 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 Δpam. Comparing to NK-101, the growth rate, asexual spore production capacity, pigment secretion capacity, mycelial biomass and secondary metabolites synthesis capacity have decreased in Δpam. Based on the above factors mentioned, we demonstrated that the PAM gene is closely related to the growth, asexual spore production, pigment secretion and secondary metabolites in NK-101.
Key words: molecular biology; Pestalotiopsis malicola; PAM gene; agrobacterium-mediated transformation; gene knockout
发表期数: 2014年3月第5期
引用格式: 颜菲,王宇,李强,等. 产紫杉醇真菌NK-101中PAM基因的功能研究[J]. 中国科技论文在线精品论文,2014,7(5):453-461.
 
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