High efficiency transformation of Tetrahymena using a codon-optimized neomycin resistance gene

被引:83
|
作者
Mochizuki, Kazufumi [1 ]
机构
[1] Austrian Acad Sci, Inst Mol Biotechnol, IMBA, A-1030 Vienna, Austria
基金
奥地利科学基金会;
关键词
Tetrahymena; neoTet; neo3; neo4; Codon usage; Biolistic transformation;
D O I
10.1016/j.gene.2008.08.007
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Tetrahymena thermophila is a useful model for the study of eukaryotic biology. A neomycin resistance gene (neo) has been developed that was optimized for the codon usage of T thermophila. Using this codon-optimized neo gene (neoTet), a new drug resistance marker cassette, neo4, has been constructed. The neo4 cassette resulted in about ten times more drug resistant transformants than a cassette containing the noncodon-optimized original neo gene. The new cassette enables transgenic Tetrahymena strains to be created with high efficiency. This study also emphasizes the importance of codon optimization in transgene expression in Tetrahymena. (c) 2008 Elsevier B.V. All rights reserved.
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页码:79 / 83
页数:5
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