Iranian Journal of Biotechnology, ( ISI ), Volume (11), No (1), Year (2013-2) , Pages (23-27)

Title : ( Identification of a Specific Pseudo attP Site for Phage phiC3 Integrase inthe Genome of Chinese Hamster in CHO-K1 Cell Line )

Authors: Mohammad Hadi Sekhavati , Kianoush Dormiani , Kamran Ghaedi , Yahya Khazaie , Morteza Hosseini , Mojtaba Tahmoorespur , Mohammadreza Nassiri , Mahboubeh Foruzanfar , Mohammad Hossein Nasr Esfahani ,

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Abstract

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.

Keywords

, attP Site, Phage phiC3 Integrase, Chinese Hamster, CHO-K1 Cell Line
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@article{paperid:1032297,
author = {Sekhavati, Mohammad Hadi and Kianoush Dormiani and Kamran Ghaedi and Yahya Khazaie and Morteza Hosseini and Tahmoorespur, Mojtaba and Nassiri, Mohammadreza and Mahboubeh Foruzanfar and Mohammad Hossein Nasr Esfahani},
title = {Identification of a Specific Pseudo attP Site for Phage phiC3 Integrase inthe Genome of Chinese Hamster in CHO-K1 Cell Line},
journal = {Iranian Journal of Biotechnology},
year = {2013},
volume = {11},
number = {1},
month = {February},
issn = {1728-3043},
pages = {23--27},
numpages = {4},
keywords = {attP Site; Phage phiC3 Integrase; Chinese Hamster; CHO-K1 Cell Line},
}

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%0 Journal Article
%T Identification of a Specific Pseudo attP Site for Phage phiC3 Integrase inthe Genome of Chinese Hamster in CHO-K1 Cell Line
%A Sekhavati, Mohammad Hadi
%A Kianoush Dormiani
%A Kamran Ghaedi
%A Yahya Khazaie
%A Morteza Hosseini
%A Tahmoorespur, Mojtaba
%A Nassiri, Mohammadreza
%A Mahboubeh Foruzanfar
%A Mohammad Hossein Nasr Esfahani
%J Iranian Journal of Biotechnology
%@ 1728-3043
%D 2013

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