Identification of a Specific Pseudo <italic>attP</italic> Site for Phage phiC3 Integrase in the Genome of Chinese Hamster in CHO-K1 Cell Line
Identification of a Specific Pseudo <italic>attP</italic> Site for Phage phiC3 Integrase in the Genome of Chinese Hamster in CHO-K1 Cell Line
عنوان فارسی :
Identification of a Specific Pseudo <italic>attP</italic> Site for Phage phiC3 Integrase in the Genome of Chinese Hamster in CHO-K1 Cell Line
عنوان انگلیسی :
Identification of a Specific Pseudo <italic>attP</italic> Site for Phage phiC3 Integrase in the Genome of Chinese Hamster in CHO-K1 Cell Line
چکیده انگلیسی:
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.
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