Author/Authors :
Hosseini Morteza نويسنده , Khazaie Yahya نويسنده , Ghaedi Kamran نويسنده , Sekhavati Mohammad Hadi نويسنده Department of Animal Science, Ferdowsi University of Mashhad, Mashhad, Iran , Tahmoorespur Mojtaba نويسنده Department of Animal Science, Ferdowsi University of Mashhad, Mashhad, Iran , Dormiani Kianoush نويسنده Department of Molecular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran , Nassiri Mohammad Reza نويسنده Department of Animal Science, Ferdowsi University of Mashhad, Mashhad, Iran , Foruzanfar Mahboubeh نويسنده Department of Molecular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran , Nasr Esfahani Mohammad Hossein نويسنده Department of Molecular Biotechnology - Cell Science Research Center - Royan Institute for Biotechnology, ACECR
Abstract :
Background: PhiC31 integrase is a DNA site-specific recombinase integrates DNA into the chromosomes between the two sites of attB and attP. Several pseudo attPs have been identified in mammalian genomes with critical features for long-term expression of transgene. In this manuscript, we report a novel intrinsic pseudo attP site named CHOL1 in the Chinese hamster genome implementing an inverse Polymerase Chain Reaction (IPCR).
Objectives:Identification of pseudo attP site(s) of Tenecteplase cDNA integration in the genome of stable transformed CHO cell line.
Materials and Methods: First, genome was extracted from a stable transformed CHO cell line expressing Tenecteplase. By creating of minicircle DNA in the last step, sequencing was performed.
Results: We obtained one band. BLAST analysis of the respective sequence of inverse PCR band identified a pseudo attP site.
Conclusions: Data demonstrated that the phiC31 integrase provides a suitable insertion site in the genome to express the gene of interest.
