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Development of a method to produce functional chimeric genes via single-strand annealing pathway§

미생물학회지 2020년 56권 4호 p.351 ~ 355
안규진, 최주희, 임예슬, 김민구, 김민구, 배성호,
소속 상세정보
안규진 ( Ahn Kyu-Jin ) - Inha University College of Natural Science Department of Biological Sciences
최주희 ( Choi Ju-Hee ) - Inha University College of Natural Science Department of Biological Sciences
임예슬 ( Lim Ye-Seul ) - Inha University College of Natural Science Department of Biological Sciences
김민구 ( Kim Min-Ku ) - Inha University College of Natural Science Department of Biological Sciences
김민구 ( Kim Min-Ku ) - Inha University College of Natural Science Department of Biological Sciences
배성호 ( Bae Sung-Ho ) - Inha University College of Natural Science Department of Biological Sciences

Abstract


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.

키워드

ASN1; ASN2; asparagine synthetase; chimeric gene; single-strand annealing

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