Here, we demonstrate the precise excision of attP- and attB-flanked DNA from the plastid genome mediated by the large serine recombinase Bxb1. We transformed the tobacco plastid genome with the pTCH-PB vector containing a stuffer fragment of DNA flanked by directly oriented nonhomologous attP and...
Before microinjection, the plasmid vector containing the donor transgene and a single cognate attB-GT site are converted into (vector- less) DNA minicircles and purified. Along with Bxb1 Int, the minicircles are delivered into zygotes carrying the attP-GT landing pad allele, enabling precision ...
摘要:本发明公开了一种通过优化提高Bxb1酶的重组效率的方法,至少包括以下步骤:(1)基于Bxb1酶结构,挑选出影响Bxb1酶活性的突变位点;(2)对步骤(1)中挑选出来的突变位点进行定向突变获得不同的Bxb1酶突变体;(3)宿主细胞中建立荧光assay,对步骤(2)中不同的Bxb1酶突变体的重组效率进行评估。本发明通过定向突变和荧...
Background The mycobacteriophage large serine recombinase Bxb1 catalyzes site-specific recombination between its corresponding attP and attB recognition sites. Previously, we and others have shown that Bxb1 has catalytic activity in various eukaryotic species including Nicotiana tabacum, Schizosaccharomyces ...
Schema for using single attP-GT site mouse strains targeted by RMKI. Before microinjection, the plasmid vector containing the donor transgene and a single cognate attB-GT site are converted into (vector-less) DNA minicircles and purified. Along with Bxb1 Int, the minicircles are delivered into...
smegmatis phage recombinase in particular the recombinases are phiRv1 or Bxb1 recombinase. The method also encompasses the use of pseudo attP and attB recombination sites as well as a third and fourth, and a fifth and sixth recombination site.PADIDAM, MALLA...
In addition, in the original version of Figure 3a, the pLN-ENR-GFP plasmid was incorrectly labeled with attB when the black box should indicate attP.doi:10.1038/nmeth0906-763Nkrumah, Louis JMuhle, Rebecca AMoura, Pedro AGhosh, Pallavi
The orientation of mycobacteriophage Bxb1 integration is solely dependent on the central dinucleotide of attP and attB. Mol Cell. 2003; 12 :1101–1111. doi: 10.1016/S1097-2765(03)00444-1. [ Cross Ref ]Ghosh P, Kim AI, Hatfull GF. The orientation of mycobacteriophage Bxb1 integration is...