文档介绍：
产紫杉醇真菌 NK-101 中 PAM 基因的功能
性研究#
颜菲,王宇,李强,马天竺,朱旭东,潘皎**
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(南开大学生命科学学院,天津 300071)
摘要: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:
)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。
关键词:拟盘多毛孢菌;PAM 基因;丝状真菌转化;基因敲除
中图分类号:Q78;
The Functional Research on PAM Gene of Taxol-Producing
Fungus NK-101
YAN Fei, WANG Yu, LI Qiang, MA Tianzhu, ZHU Xudong, PAN Jiao
(College of Life Sciences, Nankai University, TianJin 300071)
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: ) in Taxus
canadensis, we have found the homologous gene in NK-101 and named it PAM gene. Based on
the homologous bination 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
synthes