Structural analysis of the lines showed that site-specific integration occurred by recombination between the genomic attP site and the attB site of the donor plasmid. We can conclude from the results that phiC31 integrase has the ability to mediate the site-specific integration of transgenes into ...
Site-specific gene integration in cultured silkworm cells mediated by phiC31 integrase The integrase from the Streptomyces bacteriophage phiC31 carries out efficient recombination between an attP site in the phage genome and an attB site in the host chromosome. In the present study, we have used th...
Thus, there is a strict spacing requirement within attB, possibly for correct positioning of the catalytic serine relative to the scissile phosphate in the active site. Finally, using integrase S12A we confirmed the inability of attL and attR or other combinations of sites to form a stable ...
Phage PhiC31 integrase integrates attB-containing plasmid into pseudo attP site in eukaryotic genomes in a unidirectional site-specific manner and maintains robust transgene expression. Few studies, however, explore its potential in livestock. This study aims to discover the molecular basis of PhiC31 ...
In mammalian cells, phiC31 integrase can recombine its own sites and can also integrate plasmids with the attB site into native sequences that bear partial identity to attP, referred to as pseudo attP sites. The enzyme recognizes a hierarchy of such sites and, in cell lines, appears to have...
链霉菌噬菌体PhiC31整合酶是一种位点特异整合酶(Site-specific recombinase, SSR),其能够精确并单向的促进attP位点与attB位点的重组,是继Cre/Loxp、FLP之后又一个倍受关注的位点特异整合酶,该整合酶已经成为基因治疗、转基因研究等领域的常用技术手段。 技术特点:φC31 整合酶系统在促进DNA链之间位点的特异重组时,不...
利用这个原理,研究者最先在细菌、酵母、和哺乳动物细胞中实现了,在PhiC31整合酶的催化下将包含 attB位点序列的质粒整合到包含attP位点(或称为docking site,锚定位点)的基因组上。2014年Groth等人首次证明利用PhiC31整合系统,在果蝇细胞或胚胎中也能高效地实现转基因。这对于果蝇转基 因技术,无论在效率和应用范围...
A phiC31 integrase and a neo(r) gene expression plasmid containing the phiC31 attB sequence were co-delivered by electroporation into Jurkat cells. After G418 selection, integration site analyses were performed using linear amplification mediated-polymerase chain reaction (LAM-PCR). ED40515(-) cells...
Unlike Cre and Flp, phiC31 recognizes two heterotypic sites, attB and attP, for recombination. While the phiC31 system has been recently applied in mouse and human cell lines and in Drosophila , it has not been demonstrated whether it can also catalyze efficient DNA recombination in zebrafish....
Seminal studies by the Calos laboratory demonstrated that the phiC31 integrase was capable of integrating plasmid with an attB site into mammalian genomes at sites that approximated the attP site. We describe the use of attB-containing plasmids with insulated reporter genes for the successful ...