2024 Phic31 attp

Phic31 attp

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WebFeb 28, 2014 · Abstract. PhiC31 integrase-mediated gene delivery has been extensively used in gene therapy and animal transgenesis. However, random integration events are … WebMay 3, 2007 · The minimal sites for ϕC31 integrase have been defined as a 39 bp attP site and a 34 bp attB site ( 10 ). Under in vitro conditions ϕC31 integrase converts ∼80% of attB and attP to products in the absence of accessory proteins and there are no restrictions on the topology of the substrates ( 16, 27, 28 ).

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WebJul 21, 2006 · Both Bxb1 and phiC31 integrases mediate integrative recombination without a requirement for divalent cations, cofactors or DNA supercoiling, making them ideally suited for targeted gene delivery... WebphiC31 integrase: Chromosome 4 (102F4), vasa promoter, marked with GFP and RFP Chromosome 2 attP Landing Site vas-int; attP40 (Stock 13-20) Genotype: y w M (eGFP, vas-int, dmRFP)ZH-2A; P {CaryP}attP40 attP site: 2 nd chromosome (25C6), marked with yellow+ phiC31 integrase: X chromosome (2A3), vasa promoter, marked with GFP and RFP flights orlando to denver co

phiC31-integrase-mediated, site-specific ... - Kyushu University

WebJan 1, 2013 · 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... Webthrough recombination at sites with sequence similarity to the native attP site (termed pseudo-attP sites). phiC31 integrase can integrate a plasmid of any size, as a single copy, … WebIt is known that phiC31 integrase has the capacity to mediate recombination between the target sequences attP and attB. To test the availability of site-specific integration in the silkworm, we first examined the efficiency of recombination between the target sites of the two plasmids in silkworm embryos and found that the frequency of ... flights orlando to cincinnati

phiC31 stocks available in the Fly Facility Fly Facility

PhiC31 recombination system demonstrates heritable germinal

WebOct 28, 2024 · 7, when a cell containing the variant phiC31 integrase that recognizes the pseudo-attP site, a recombination between the pseudo-phiC31 attP site and the phiC31 attB site occurs and the orientation of the CMV promoter in the selection construct is reversed. The reversed CMV promoter then drives the expression of the GFP reporter gene. Web本发明涉及一种方法，具体是一种制备包含编码多模块生物合成酶的多个基因在内的基因簇构建体的方法，其包括以下工序：(A)制备可重构基因簇构建体并具有可交替连接结构的多个DNA片段的工序；(B)通过将工序(A)中制备的多个DNA片段在溶液中混合来交替连接多个DNA片段，从而得到基因簇构建体的 ... cherry stoner toolWebExpression of sgRNA under D. melanogaster U6-3 _(CR31539) promoter & delivery of D.mel-Actin5C::EGFP-2A-puro sgRNA cassette through PhiC31 RCME to AttP acceptor cell line/organism Depositor Norbert Perrimon flights orlando to columbus ga

"WebphiC31 attP Alt name phiC31 phage attachment site Species Streptomyces phage phiC31 Insert Size (bp) 221 Cloning Information Cloning method Restriction Enzyme 5′ … " - Phic31 attp

Phic31 attp

WebSep 14, 2012 · The target sequence involves a reporter gene encoding green fluorescent protein (GFP), which is flanked by the attB and attP recognition sites for the phiC31 integrase. This sequence disruptively separates a gusA coding sequence from an upstream rice actin promoter. We succeeded in producing site-specific recombination events in the … WebOct 20, 2013 · A-E. Assays φC31 integrase activity on attachment sites in different configurations. The indicated plasmids containing either attB or attP sites were incubated in the absence or in the presence of 0 nM (lane 1), 100 nM (lane 2), 200 nM (lane 3), 350 nM (lane 4) and 700 nM (lane 5) φC31 integrase.

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WebFeb 15, 2024 · The attP site (5'-GTGCCCCAACTGGGGTAACCTttGAGTTCTCTCAGTTGGGGG-3') is one of the two target sites of the phiC31:int integrase. phiC31:int mediates recombination between two non-identical target sites, attP and attB, each of which contains a short integration core flanked by imperfect inverted repeats.

WebMar 21, 2012 · The Bxb1 recombinase is a 500 amino acid protein that binds minimal recognition sites attP and attB that are 39 bp and 34 bp, respectively and enzymatically executes uni-directional site-specific recombination [ 9 ]. WebIn addition, combining PhiC31 integrase with R4 integrase allows for isogenic expression from a defined genomic locus, providing the ideal solution for comparative analysis of gene families, isoforms or orthologs. ... that express your gene of interest by having you initially generate a retargetable platform cell line containing the R4 attP ...

WebJun 17, 2010 · 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. WebMar 23, 2024 · 该研究一个重要的创新点是通过改造酵母双杂交系统的载体质粒(Yang et al., 2024), 将诱饵蛋白所在载体(pGBKT7)插入ATTB位点, 将捕获蛋白所在载体(pGADT7)插入ATTP位点, 分别在2个载体上插入来自噬箘体PhiC31的表达盒。利用PhiC31介导的ATTB位点和ATTP位点发生重组, 使互作的2 ...

WebSet up embryo collection cages with germline-supplied phiC31 females and males that carry the Cas9-mediated attP knock-in. Embryos collected from these cages are used for fly transformation. Follow Subheading 3.2 to generate transgenic flies with the integrated imaging tags. Negative selection, i.e., loss of the marker introduced with the Cas9 ...

WebThe phiC31 integrase catalyzes unidirectional recombination between two attachment sites, attP and attB, leading to the formation of attL and attR sites . A circular construct harboring an attB site can be efficiently and specifically integrated into an attP site located on the genome . The phiC31 ... cherry stone removerWebphiC31 integrase is a site-specific recombinase from a bacteriophage that has become a useful tool in mammalian cells. The enzyme normally performs precise, unidirectional … flights orlando to clevelandWebPhiC31 integrase, a site-specific large serine recombinase, is a useful tool for genome engineering in a variety of eukaryotic species and cell types. PhiC31 integrase performs efficient recombination between its attB site and either its own placed attP site or a partially mismatched genomic pseudo attP site. flights orlando to el paso texasWebPhiC31 integrase [ edit] Without much doubt, Ser integrases are the current tools of choice for integrating transgenes into a restricted number of well-understood genomic acceptor sites that mostly (but not always) mimic the phage att P site in that they attract an att B-containing donor vector. flights orlando to dominican republicWebFeb 15, 2024 · The attB site (5'-TGCGGGTGCCAGGGCGTGCCCttGGGCTCCCCGGGCGCGTACTCC-3') is one of the two target sites of the phiC31:int integrase. phiC31:int mediates recombination between two non-identical target sites, attP and attB, each of which contains a short integration core … flights orlando to gatlinburg tnWebPhiC31 Integrase Expression Plasmid Put the integrase into your PhiC31 and PinPoint genome editing projects with this vector that expresses PhiC31 Integrase for one-step … flights orlando to flagstaffWebJun 12, 2024 · The phage-derived phiC31 integrase is a useful tool for mediating sequence-specific genomic integration in mammalian cells, recombining donor plasmids bearing the attB recognition site with introduced genomic attP sites or endogeneous pseudo-attP sites having partial identity to attP. flights orlando to destin