identification of a specific pseudo attp site for phage phic3 integrase in the genome of chinese hamster in cho-k1 cell line

Authors

mohammad hadi sekhavati

kianoush dormiani

kamran ghaedi

yahya khazaie

morteza hosseini

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.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

Identification of a Specific Pseudo attP Site for Phage phiC3 Integrase in the Genome of Chinese Hamster in CHO-K1 Cell Line

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 Chi...

full text

Identification of a Specific Pseudo attP Site for Phage PhiC31 Integrase in Bovine Genome

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...

full text

identification of a specific pseudo attp site for phage phic31 integrase in bovine genome

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: ...

full text

creation of tenecteplase-producing cho cell line using site-specific integrase from the phage φc31

background: the aim of this study was to produce a stable cho cell line expressing tenecteplase. materials and methods: in the first step, the tenecteplase coding sequence was cloned in a pdb2 vector containing attb recognition sites for the phage φc31 integrase. then, using lipofection, the cho cells were co-transfected with constructed recombinant plasmid encoding tenecteplase and attb recogn...

full text

study of cohesive devices in the textbook of english for the students of apsychology by rastegarpour

this study investigates the cohesive devices used in the textbook of english for the students of psychology. the research questions and hypotheses in the present study are based on what frequency and distribution of grammatical and lexical cohesive devices are. then, to answer the questions all grammatical and lexical cohesive devices in reading comprehension passages from 6 units of 21units th...

Propranolol induced chromosomal aberrations in Chinese hamster ovary cell line

Propranolol (PL), a non-selective beta-blocker, is a cardiovascular drug widely used to treat hypertension. The present study was concerned with assessing the cytogenetic effects of this drug on Chinese hamster ovary (CHO) cell line. MTT assay was then carried out to determine the cytotoxicity index (IC50) of the drug. The IC50 value of PL was 0.43±0.02 mM. To investigate the clastogenic effect...

full text

My Resources

Save resource for easier access later


Journal title:
iranian journal of biotechnology

Publisher: national institute of genetic engineering and biotechnology

ISSN 1728-3043

volume 11

issue 1 2013

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023