Recombination of attP and attB sites creates attL and attR sites. Gateway vectors contain modified versions of the att sites so that scientists can easily clone in their desired DNA sequences. Gateway technology relies on the two reactions described below:

Gateway cloning is a highly efficient alternative to restriction cloning and does not require the use of restriction enzymes. Instead, an insert is moved into a vector through a two-step recombination process that takes advantage of integration and excision …

The gene of interest must be flanked by the appropriate att sites, either attL (100 bp) in an Entry clone or attB (25 bp) in a PCR product. For Entry clones, everything between the attL sites will be shuttled into the Gateway® Destination vector containing attR sites, and a PCR product flanked by attB sites must be shuttled into an attP ...

2017年7月14日 · We tested attB HK sites of three different lengths to avoid potential interference with bla function and protein export, 51 bp (violet), 33 bp (teal), and 23 bp (black). To increase the number of potential open reading frames, we introduced a T A nucleotide change into the attB sequence, indicated in red.

Serine integrases catalyze the integration of bacteriophage DNA into a host genome by site-specific recombination between 'attachment sites' in the phage (attP) and the host (attB). The reaction is highly directional; the reverse excision reaction …

(A) List of attB sites; attB0 is the E. coli bacteriophage attachment site (Landy 1989). Shaded area indicates 7-bp overlap (Int cut sites); underlined bases indicate changes from attB0.

2025年1月11日 · In molecular cloning, attP and attB sites are incorporated into vectors and host genomes to insert foreign DNA. Systems like Gateway cloning exploit these sites’ natural recombination mechanisms, enabling efficient and accurate genetic material insertion or removal.

2022年7月20日 · Attachment sites attP and attB are bound by Bxb1 dimers, followed by synapsis through intermolecular binding of coiled-coil motifs (CC, pink). DNA sequences are then cleaved in the middle by...

In-vitro Gateway cloning models the recombination reactions that happen when a lambda phage infects a bacterium in-vivo. A lambda phage contains a region of DNA called an attP site. When the phage infects a bacterium, this attP site comes into contact with a region called the attB site in the bacterial genome.

The first step in the Gateway cloning process is to amplify the target sequence with primers including so-called attB sites. In the CLC Genomics Workbench, you can add attB sites to a sequence fragment in this way: Toolbox in the Menu Bar | Molecular Biology Tools | Cloning and Restriction Sites ()| Gateway Cloning | Add attB Sites ()