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A Profile of Native Integration sites Used by φC31 Integrase in the Bovine Genome

Journal of Genetics and Genomics 2012年第5期39卷 217-224页

作者： Lijuan Qu Qingwen Ma Zaiwei Zhou Haiyan Ma Ying Huang Shuzhen Huang Fanyi Zeng Yitao Zeng Shanghai Institute of Medical Genetics Children＇s Hospital of Shanghai Shanghai Jiao Tong University School of Medicine 24/1400 West Beijing Road Shanghai 200040 China Key Lab of Medical Embryo Molecular Biology Ministry of Health Shanghai 200040 China Shanghai Key Lab of Embryo and Reproduction Engineering Shanghai Municipal Committee of Science and Technology Shanghai 200040 China Institute of Medical Science Shanghai Jiao Tong University School of Medicine 280 South Chongqing Road Shanghai 200025 China

The Streptomyces phage φC31 integrase can efficiently target attB-bearing transgenes to endogenous pseudo attp sites within mammalian genomes. To better understand the activity of φC31 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 φC31-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 specific integrase system has great potential for applied modifications in the bovine genome were analyzed. We conclude that this site- of the bovine genome.

关键词： φC31 integrase pseudo attp sites Sequence motif Repetitive elements Bovine genome

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