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KMID : 0364820200560040351
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Korean Journal of Microbiology
2020 Volume.56 No. 4 p.351 ~ p.355
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Development of a method to produce functional chimeric genes via single-strand annealing pathway§
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Ahn Kyu-Jin
Choi Ju-Hee
Lim Ye-Seul
Kim Min-Ku
Kim Min-Ku
Bae Sung-Ho
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Abstract
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Duplicated genes with similar sequences are commonly observed in eukaryotic genomes, some of which show sufficient homology to allow for recombination. ASN1 and ASN2 are an example of duplicated genes that share highly homologous sequence throughout the open reading frame. In this study, we devised a genetic method to produce functional chimeric ASN genes via single-strand annealing pathway. Creation of a DNA doublestrand break between ASN1 and ASN2 using HO endonuclease generated mutant cells with chimeric genes at significantly high frequency. All mutant cells exhibited normal growth in the medium lacking asparagine, suggesting that the chimeric genes are functional. Sequence analysis of chimeric genes revealed that they had different single junctions. Our results provide a method to produce a variety of chimeric genes simultaneously by randomly fusing homologous genes.
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KEYWORD
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ASN1, ASN2, asparagine synthetase, chimeric gene, single-strand annealing
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