Functional expression of the Cre recombinase in actinomycetes

被引:49
|
作者
Fedoryshyn, Marta [1 ]
Welle, Elisabeth [1 ]
Bechthold, Andreas [1 ]
Luzhetskyy, Andriy [1 ]
机构
[1] Univ Freiburg, Inst Pharmazeut Wissensch, D-79104 Freiburg, Germany
关键词
site-specific recombinase; Cre/loxP; actinomycetes; natural products;
D O I
10.1007/s00253-008-1382-9
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Site-specific recombinases revolutionized "in vivo" genetic engineering because they can catalyze precise excisions, integrations, inversions, or translocations of DNA between their distinct recognition target sites. We have constructed a synthetic gene encoding Cre recombinase with the GC content 67.7% optimized for expression in high-GC bacteria and demonstrated this gene to be functional in Streptomyces lividans. Using the synthetic cre(a) gene, we have removed an apramycin resistance gene flanked by loxP sites from the chromosome of S. lividans with 100% efficiency. Sequencing of the chromosomal DNA part showed that excision of the apramycin cassette by Cre recombinase was specific.
引用
收藏
页码:1065 / 1070
页数:6
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