Seamless deletion of a large DNA fragment in the taxol-producing fungus Pestalotiopsis microspora

被引：2
|
作者：
Chen, Longfei [1 ,2 ]
Li, Yingying [1 ,2 ]
Zhang, Qian [1 ,2 ]
Akhberdi, Oren [1 ,2 ]
Wei, Dongsheng [1 ,2 ]
Pan, Jiao [1 ,2 ]
Zhu, Xudong [1 ,2 ,3 ]
机构：
[1] Nankai Univ, Coll Life Sci, State Key Program Microbiol, Tianjin 300071, Peoples R China
[2] Nankai Univ, Coll Life Sci, Dept Microbiol, Tianjin 300071, Peoples R China
[3] Beijing Normal Univ, Beijing Key Lab Gene Engn Drug & Biotechnol, Inst Biochem & Biotechnol, Coll Life Sci, Beijing 100875, Peoples R China
来源：
MYCOSCIENCE | 2017年 / 58卷 / 01期
基金：
中国国家自然科学基金;
关键词：
ATMT; 5-FOA; Counter selection; Orotidine 5 '-phosphate decarboxylase; Seamless genome reduction; GENE DISRUPTION; INSERTIONAL MUTAGENESIS; MEDIATED TRANSFORMATION; EFFICIENT TOOL; CHEMISTRY; MARKER;
D O I：
10.1016/j.myc.2016.08.003
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
To reduce the unnecessary gene clusters in the taxol-producing fungus Pestalotiopsis microspora, we report the development of an effective DNA deletion method that relies on a deletion cassette constructed with the Gateway-technique and overlap extension PCR, using the orotidine 5'-phosphate decarboxylase as recyclable marker for selection. By this approach, two adjacent DNA sequences can be sequentially deleted in a single transformation mediated by Agrobacterium tumefaciens, resulting in the deletion of a large DNA fragment. Additionally, the selection marker is spontaneously eliminated in this process. We used this method to successfully remove the mus53 locus of P. microspora. (C) 2016 The Mycological Society of Japan. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码：35 / 39
页数：5
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