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작성자 Luella
댓글 0건 조회 7회 작성일 25-02-08 20:14

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The specificity profile for Cre reveals its relative preference for the canonical base at every position within the loxP site. We sought to develop a system for profiling recombinase specificity via identification of bona fide recombinase substrates from an enormous in vitro library of possible targets. Recombinase specificity profiling of wild-kind Cre. As well as, Rec-seq recognized a contribution from a secondary residue previously unknown to participate in specifying positions 5-7. Ala substitution at Gln94 resulted in decrease specificity at positions 6 and 7 but compensatory increases elsewhere (Fig. 2e), regardless that Gln94 does in a roundabout way contact the DNA, however as a substitute engages in hydrogen bonds with Gln9035. Together, Rec-seq profiling clarifies the numerous interactions that collectively define Cre recognition at positions 5-7, and highlights the important roles of secondary and oblique interactions. These lengthy-range results cannot be inferred from the Cre:loxP construction, highlighting the utility of unbiased, high-decision specificity profiling. Determinants of Cre:loxP specificity recognized by Rec-seq on wild-type Cre and Ala-substituted Cre variants

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