![]() ![]() SSR between attP/I and attB sites is mediated by an integrase (Int) and a recombination directionality factor (RDF). The final expression vectors can also be used for releasing the attR cassette for constructing new vectors. Site-specic recombination (SSR) systems are employed for transfer of mobile genetic elements (MGEs), such as lysogenic phages and integrative conjugative elements (ICEs). The enzyme efficiently mediates recombination between two distinct sequences, attP and attB, producing recombinant product sites, attL and attR. Recombination between attB site and the human locus Xq22.1 This recombination generates a pattR which has been isolated by inverse PCR. With the increasing number of recombination vectors constructed with this protocol, the likeliness of releasing the attB cassette from an existing vector, rather than synthesizing it with PCR, will increase. The intermediate vectors generated in this study can be used for releasing the attB cassette to convert other vectors using the same protocol described here. An attB cassette flanked by several restriction enzyme sites was inserted in a cloning vector, and then subcloned into existing vectors to be converted to construct intermediate vectors containing the attB cassette, which were then converted to recombination expression vectors by in vitro recombination. The reaction is performed between an attB-flanked DNA fragment and. In this report, we describe the conversion of a set of widely used conventional vectors to Gateway recombination expression vectors. cerevisiae relies on homologous recombination for its maintenance (Contamine and. Based on these important observations, we employed both Agrobacterium-mediated insertional mutagenesis and bioinformatics to. Thus, it is desirable to convert existing conventional vectors to recombination vectors. In recent years, restriction-less recombination cloning systems based on site-specific recombinase with high efficiency have been proven to be very successful. ![]()
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