identification of a specific pseudo attp site for phage phic3 integrase in the genome of chinese hamster in cho-k1 cell line
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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.
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Journal title:
iranian journal of biotechnologyPublisher: national institute of genetic engineering and biotechnology
ISSN 1728-3043
volume 11
issue 1 2013
Keywords
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