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PubMed · 2699239

An efficient and site-specific gene trimming method.

H M Eun·Y Kang·S M Kang·Y S Bae·J W Yoon
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Abstract

Trimming a DNA strand into a precisely determined fragment can be carried out efficiently by an improved method involving a site-specific trim-primer and a single-stranded DNA template which is generated from a multifunctional vector, pTZ18R, and linearized by using an Eco RI-pTZ18R splinter. A complementary DNA strand is synthesized by DNA polymerase I (Klenow), and the 3'-end of the template upstream from the annealed primer is trimmed by subsequent T4 DNA polymerase reaction. An ATG translation initiator codon or a termination codon can be incorporated into the trim-primer, providing versatility to this single-stranded DNA-initiated gene trimming method that can be applied to subcloning and expression of any DNA fragment with known terminal sequences.

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BiotechnologyCloning, MolecularDNADNA Polymerase IDNA RepairDNA, Single-StrandedGenetic TechniquesHumansInsulinProinsulinProtein PrecursorsJournal ArticleResearch Support, Non-U.S. Gov't

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H M Eun, Y Kang, S M Kang, Y S Bae, J W Yoon. 1989. An efficient and site-specific gene trimming method.. https://pubmed.ncbi.nlm.nih.gov/2699239/

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