文献类型：期刊文献

英文题名：PiggyBac transposon-mediated mutagenesis and application in yeast Komagataella phaffii

作者：Zhu, Jinxiang[1];Zhu, Qiaoyun[1];Gong, Ruiqing[1];Xu, Qin[1];Cai, Menghao[1];Jiang, Tianyi[3];Zhou, Xiangshan[1];Zhou, Mian[1];Zhang, Yuanxing[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]SCICB, Shanghai 200237, Peoples R China;[3]Roche R&D Ctr China Ltd, Shanghai, Peoples R China

年份：2018

卷号：40

期号：9-10

起止页码：1365

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;EI(收录号：20182905550031);WOS:【SCI-EXPANDED(收录号:WOS:000444565700009)】；

基金：This work was sponsored by National Natural Science Foundation of China (31600056); the Fundamental Research Funds for the central Universities (222201714021); Shanghai Chenguang Program (15CG27).

语种：英文

外文关键词：Mutagenesis; Komagataella phaffii; piggyBac; Transposon

摘要：ObjectiveAround one-fourth of the Komagataella phaffii genes encode hypothetical proteins with unknown functions. However, lack of powerful tools for genetic screening in K. phaffii significantly limits the functional analysis of these unknown genes. Transposon mutagenesis has been utilized as an insertional mutagenesis tool in many other organisms and would be extremely valuable if it could be applied in K. phaffii.ResultsIn this study, we investigated in K. phaffii the transposition activity and efficiency of piggyBac (PB) transposon, a DNA transposon from the cabbage looper moth Trichoplusia ni through the integrated-plasmid system. We also designed a binary-plasmid system which could generate stable mutants. Finally we evaluated the quality of this mutagenesis system by a simple screening for functional genes involved in K. phaffii carbon catabolite repression.ConclusionsOur results demonstrate that PB-mediated mutagenesis could be a feasible and useful tool for functional gene screening in K. phaffii.