In the second round, the BAC to be added was linearised with the very rare-cutting enzyme I-PpoI and electroporated into recombination efficient E元50 bacteria carrying the other BAC. In the first round, the inserts of the two overlapping BACs were subcloned into modified BAC vectors using homologous recombination. Two rounds of homologous recombination were carried out in the E元50 strain of bacteria which can be induced for the Red genes. Results The method was used to link a 61-kb insert carrying the final 5 exons of the human CFTR gene onto a 160-kb BAC carrying the first 22 exons. Here we describe a system for linking two or more overlapping BACs into a single clone by homologous recombination. However, large genes often span more than one BAC, and single BACs covering the entire region of interest are not available. Sequenced BACs are available for most of the human and mouse genomes and in many cases these contain intact genes. Recombining overlapping BACs into a single larger BACĭirectory of Open Access Journals (Sweden)įull Text Available Abstract Background BAC clones containing entire mammalian genes including all the transcribed region and long range controlling elements are very useful for functional analysis.
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June 2023
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