Creating recombinant plasmids

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Question: Ligation of the multiple ? fragments and pBLU produces a number of different hybrid molecules, although not as many as you might expect. Each clonable fragment must have one BamHI and one HindIII end, to combine with the BamHI and HindIII ends of the pBLU backbone and form a circular recombinant plasmid. Only 5 of the 13 BamHI/HindIII framgnets from the linear ? genome satisfy this requirement: 493 bp, 784 bp, 2,396 bp, 2,409 bp, and 4,148 bp. A sixth clonable fragment of 9,866 bp is derived from the COS circle. Multiple copies of a single fragment or combinations of two different ? fragments can also be ligated into a single pBLU backbone. Three fragment plasmids can be constructed by combining the pBLU backbone with one of hte five fragments having one BamHI end and one HindIII end and with one of the eight fragments having two BamHI or two HindIII ends. Constructs with more than one unit of pBLU are also formed. Oly constructs with at least one unit of pBLU backbone contain an origin necessary for replication and the ampicillin-resistance gene required for growth on selective media.

1. Make scale drawings of all potential two-fragment (vector + insert) recombinant plasmids. Include the total bp size, fragment bp sizes, and the locations of BamHI sites and HindIII sites, the origin, the amplicillin-resistance gene, and the lacZ gene.


Reference no: EM1387491

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