TY - JOUR ID - 7571 TI - Identification of a Specific Pseudo attP Site for Phage PhiC31 Integrase in Bovine Genome JO - Iranian Journal of Biotechnology JA - IJB LA - en SN - 1728-3043 AU - Sekhavati, Mohammad Hadi AU - Tahmoorespur, Mojtaba AU - Jafarpour, Farnoosh AU - Dormiani, Kianoush AU - Khazaie, Yahya AU - Ghaedi, Kamran AU - Hosseini, Sayyed Morteza AU - Forouzanfar, Mahboobeh AU - Nasr Esfahani, Mohammad Hossein AD - Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran AD - Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, I.R. IRAN AD - Department of Reproduction and Development at Reproductive Biomedicine Center, Royan Institute for Biotechnology, ACECR, Isfahan, I.R. IRAN AD - Department of Molecular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, I.R. IRAN AD - Department of Biology, School of Sciences, University of Isfahan & Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology Y1 - 2014 PY - 2014 VL - 12 IS - 4 SP - 1 EP - 9 KW - PhiC31 intagrase KW - Pseudo attP site KW - Protein expression KW - Recombination DO - 10.15171/ijb.1013 N2 - Background: PhiC31 integrase system provides a new platform in various felid of research, mainly in gene therapy and creation of transgenic animals. This system enables integration of exogenous DNA into preferred locations in mammalian genomes, which results in robust, long-term expression of the integrated transgene. Objectives: Identification of a novel pseudo attP site. Materials and Methods: Genomic DNA was extracted from primary bovine fetal fibroblast cells, which were stably transfected with EGFP and phiC31 integrase cDNAs carrying vectors. An inverse PCR was carried out for production of mini-circle DNAs and followed by sequencing. Results: A new specific pseudo attP site termed BF5 was identified in bovine genome.  This site is located in an intergenic AT rich region on chromosome 5 with similar features of other mammalian attP pseudo sites.  Furthermore, direct sequencing of generated attL site confirmed that site-specific transgene recombination was occurred at this site. Conclusions: This finding confirmed that phiC31 integrase could be feasible for production of transgenic animals for biotechnological applications. UR - https://www.ijbiotech.com/article_7571.html L1 - https://www.ijbiotech.com/article_7571_2aee096c390db0e197ea0b5e1bef2f00.pdf ER -