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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 in... Full description

Journal Title: Iranian Journal of Biotechnology 12/2014, Vol.12(4), pp.1-9
Main Author: Sekhavati, Mohammad Hadi
Other Authors: Tahmoorespur, Mojtaba , Jafarpour, Farnoosh , Dormiani, Kianoush , Khazaie, Yahya , Ghaedi, Kamran , Hosseini, Sayyed Morteza , Forouzanfar, Mahboobeh , Nasr Esfahani, Mohammad Hossein
Format: Electronic Article Electronic Article
Language: English
Subjects:
DNA
Pcr
Bos
ID: ISSN: 1728-3043 ; E-ISSN: 2322-2921 ; DOI: http://dx.doi.org/10.15171/ijb.1013
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recordid: crossref10.15171/ijb.1013
title: Identification of a Specific Pseudo attP Site for Phage PhiC31 Integrase in Bovine Genome
format: Article
creator:
  • Sekhavati, Mohammad Hadi
  • Tahmoorespur, Mojtaba
  • Jafarpour, Farnoosh
  • Dormiani, Kianoush
  • Khazaie, Yahya
  • Ghaedi, Kamran
  • Hosseini, Sayyed Morteza
  • Forouzanfar, Mahboobeh
  • Nasr Esfahani, Mohammad Hossein
subjects:
  • Bacteriophages
  • Biotechnology
  • Chromosomes
  • Complementary DNA
  • DNA
  • DNA Sequencing
  • Fetus
  • Fibroblasts
  • Gene Expression
  • Gene Therapy
  • Genes
  • Genetically Engineered Organisms
  • Genomes
  • Genomics
  • Intergenic DNA
  • Nucleotide Sequences
  • Polymerase Chain Reaction
  • Recombination
  • Transfection
  • Transgenic Animals
  • Cdna
  • Deoxyribonucleic Acid
  • DNA Sequences
  • Foetus
  • Genetic Recombination
  • Genetically Engineered Animals
  • Genetically Modified Animals
  • Genetically Modified Organisms
  • Geos
  • Gmos
  • Intergenic Spacers
  • Nucleotide Sequence Analysis
  • Nucleotide Sequencing
  • Pcr
  • Phages
  • Spacer DNA
  • Transgenic Organisms
  • Cattle
  • Viruses
  • Bos
  • Bovidae
  • Ruminants
  • Artiodactyla
  • Mammals
  • Vertebrates
  • Chordata
  • Animals
  • Eukaryotes
ispartof: Iranian Journal of Biotechnology, 12/2014, Vol.12(4), pp.1-9
description: 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: Genomic DNA was extracted from primary bovine fetal fibroblast cells, which were stably transfected with EGFP and phiC31 integrase cDNAs carrying vectors. An inverse PCR was carried out for production of mini-circle DNAs and followed by sequencing. Results: A new specific pseudo attP site termed BF5 was identified in bovine genome. This site is located in an intergenic AT rich region on chromosome 5 with similar features of other mammalian attP pseudo sites. Furthermore, direct sequencing of generated attL site confirmed that site-specific transgene recombination was occurred at this site. Conclusions: This finding confirmed that phiC31 integrase could be feasible for production of transgenic animals for biotechnological applications.
language: eng
source:
identifier: ISSN: 1728-3043 ; E-ISSN: 2322-2921 ; DOI: http://dx.doi.org/10.15171/ijb.1013
fulltext: fulltext
issn:
  • 17283043
  • 1728-3043
  • 23222921
  • 2322-2921
url: Link


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titleIdentification of a Specific Pseudo attP Site for Phage PhiC31 Integrase in Bovine Genome
creatorSekhavati, Mohammad Hadi ; Tahmoorespur, Mojtaba ; Jafarpour, Farnoosh ; Dormiani, Kianoush ; Khazaie, Yahya ; Ghaedi, Kamran ; Hosseini, Sayyed Morteza ; Forouzanfar, Mahboobeh ; Nasr Esfahani, Mohammad Hossein
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descriptionBackground: 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: Genomic DNA was extracted from primary bovine fetal fibroblast cells, which were stably transfected with EGFP and phiC31 integrase cDNAs carrying vectors. An inverse PCR was carried out for production of mini-circle DNAs and followed by sequencing. Results: A new specific pseudo attP site termed BF5 was identified in bovine genome. This site is located in an intergenic AT rich region on chromosome 5 with similar features of other mammalian attP pseudo sites. Furthermore, direct sequencing of generated attL site confirmed that site-specific transgene recombination was occurred at this site. Conclusions: This finding confirmed that phiC31 integrase could be feasible for production of transgenic animals for biotechnological applications.
subjectBacteriophages ; Biotechnology ; Chromosomes ; Complementary DNA ; DNA ; DNA Sequencing ; Fetus ; Fibroblasts ; Gene Expression ; Gene Therapy ; Genes ; Genetically Engineered Organisms ; Genomes ; Genomics ; Intergenic DNA ; Nucleotide Sequences ; Polymerase Chain Reaction ; Recombination ; Transfection ; Transgenic Animals ; Cdna ; Deoxyribonucleic Acid ; DNA Sequences ; Foetus ; Genetic Recombination ; Genetically Engineered Animals ; Genetically Modified Animals ; Genetically Modified Organisms ; Geos ; Gmos ; Intergenic Spacers ; Nucleotide Sequence Analysis ; Nucleotide Sequencing ; Pcr ; Phages ; Spacer DNA ; Transgenic Organisms ; Cattle ; Viruses ; Bos ; Bovidae ; Ruminants ; Artiodactyla ; Mammals ; Vertebrates ; Chordata ; Animals ; Eukaryotes;
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