Disruption of lig4 improves gene targeting efficiency in the oleaginous fungus Mortierella alpina 1S-4
Disruption of lig4 improves gene targeting efficiency in the oleaginous fungus Mortierella alpina 1S-4
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DOI：
10.1016/j.jbiotec.2015.05.020
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发表时间：
2015-08-20
影响因子：
4.1
通讯作者：
Ogawa, Jun
中科院分区：
文献类型：
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作者：
Kikukawa, Hiroshi;Sakuradani, Eiji;Ogawa, Jun
已知产油放线菌高山被孢霉1 S-4积累有益的多不饱和脂肪酸。我们鉴定了编码DNA连接酶4同源物的lig 4基因，其功能是通过非同源末端连接来修复双链断裂。为了提高M.阿尔卑斯山。分枝alpina 1 S-4 Delta lig 4菌株在营养生长、孢子形成和脂肪酸产生方面没有表现出缺陷，但对甲磺酸甲酯（一种引起DNA双链断裂的试剂）表现出高敏感性。重要的是，在67%的Delta lig 4菌株中发生了CBXB标记物对ura 5标记物的基因替换，并且基因靶向效率比在M中破坏lig 4基因时观察到的高21倍。alpina 1 S-4宿主菌株。Delta lig 4菌株的进一步代谢工程有望产生产生更高水平的稀有和有益的多不饱和脂肪酸的菌株，并有助于对放线菌的基础研究。(C)2015 Elsevier B. V.版权所有。
The oil-producing zygomycete Mortierella alpina 1S-4 is known to accumulate beneficial polyunsaturated fatty acids. We identified the lig4 gene that encodes for a DNA ligase 4 homolog, which functions to repair double strand breaks by non-homologous end joining. We disrupted the lig4 gene to improve the gene targeting efficiency in M. alpina. The M. alpina 1S-4 Delta lig4 strains showed no defect in vegetative growth, formation of spores, and fatty acid production, but exhibited high sensitivity to methyl methansulfonate, an agent that causes DNA double-strand breaks. Importantly, gene replacement of ura5 marker by CBXB marker occurred in 67% of Delta lig4 strains and the gene targeting efficiency was 21-fold greater than that observed in disruption of the lig4 gene in the M. alpina 1S-4 host strain. Further metabolic engineering of the Delta lig4 strains is expected to result in strains that produce higher levels of rare and beneficial polyunsaturated fatty acids and contribute to basic research on the zygomycete. (C) 2015 Elsevier B.V. All rights reserved.