A Profile of Native Integration Sites Used by 蠁C31 Integrase in the Bovine Genome

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• 作者：Lijuan Qu^a ; Qingwen Ma^a ; ^b ; ^c ; Zaiwei Zhou^a ; Haiyan Ma^a ; ^b ; ^c ; Ying Huang^a ; ^b ; ^c ; Shuzhen Huang^a ; ^b ; ^c ; Fanyi Zeng^a ; ^b ; ^c ; ^d ; ^{fzeng@sjtu.edu.cn} ; Yitao Zeng^a ; ^b ; ^c ; ^{ytzeng@stn.sh.cn}
• 关键词：蠁C31 integrase ; Pseudo attP sites ; Sequence motif ; Repetitive elements ; Bovine genome
• 刊名：Journal of Genetics and Genomics
• 出版年：2012
• 期刊代码：23_16738527
• 类别：bio
• 出版时间：20 May, 2012
• 卷：39
• 期：5
• 页码：217-224
• 文件大小：1033 K

摘要

The Streptomyces phage 蠁C31 integrase can efficiently target attB-bearing transgenes to endogenous pseudo attP sites within mammalian genomes. To better understand the activity of 蠁C31 integrase in the bovine genome, DNA sequences of 44 integration events were analyzed, and 32 pseudo attP sites were identified. The majority of these sites share a sequence motif that contains inverted repeats and has similarities to wild-type attP site. Genomic DNA flanking these sites typically contained repetitive sequence elements, such as short and long interspersed repetitive elements. These sequence features indicate that DNA sequence recognition plays an important role in guiding 蠁C31-mediated site-specific integration. In addition, BF27 integration hotspot sites were identified in the bovine genome, which accounted for 13.6%of all isolated integration events and mapped to an intron of the deleted in liver cancer 1 (DLC1) gene. Also we found that the pseudo attP sites in the bovine genome had other features in common with those in the human genome. This study represents the first time that the sequence features of pseudo attP sites in the bovine genome were analyzed. We conclude that this site-specific integrase system has great potential for applied modifications of the bovine genome.