Functional expression of the Cre recombinase in actinomycetes
Functional expression of the Cre recombinase in actinomycetes
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DOI：
10.1007/s00253-008-1382-9
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发表时间：
2008-04-01
影响因子：
5
通讯作者：
Luzhetskyy, Andriy
中科院分区：
文献类型：
--
作者：
Fedoryshyn, Marta;Welle, Elisabeth;Luzhetskyy, Andriy
位点特异性重组酶彻底改变了“体内”遗传工程，因为它们可以催化DNA在其不同识别靶位点之间的精确切除、整合、倒位或易位。我们已经构建了一个合成的基因编码Cre重组酶的GC含量为67.7%，优化高GC细菌中的表达，并证明该基因是在变铅青链霉菌的功能。利用人工合成的cre（a）基因，我们从S. 100%的效率。染色体DNA部分的测序显示，Cre重组酶对安普霉素盒的切除是特异性的。
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.