publication . Article . 2002

A plasmid vector with positive selection and directional cloning based on a conditionally lethal gene

Sergey A. Yazynin; Sergey M. Deyev; Milan Jucovi; Robert W. Hartley;
Open Access
  • Published: 25 Jul 2002 Journal: Gene, volume 169, pages 131-132 (issn: 0378-1119, Copyright policy)
  • Publisher: Elsevier BV
Abstract
Abstract A plasmid vector with a multiple cloning site (MCS) for positive selection of cloned inserts in Escherichia coli (Ec) has been devised, based on the expression plasmid (pMT416) for the bacterial ribonuclease barnase (Barn). The host is protected from the lethal effect of moderate expression of barn by expression of the gene bars, encoding its inhibitor, barstar (Bars), placed on the same plasmid. Full expression, however, is lethal. Induction is also lethal with the derived plasmid, pMT440, which has the pUC19 MCS inserted into barn. Under inducing conditions, transformation by the vector is lethal unless the product of the modified barn is inactivated ...
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Medical Subject Headings: psychological phenomena and processesanimal diseases
free text keywords: Barnase, biology.protein, biology, T-DNA Binary system, Molecular biology, Barstar, pUC19, Plasmid, Expression vector, Multiple cloning site, Cloning vector
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