Isolation of dominant negative mutants and inhibitory antisense RNA sequences by expression selection of random DNA fragments

Tatyana A. Holzmayer, Dmitry G. Pestov, Igor B. Roninson

Research output: Contribution to journalArticlepeer-review

48 Scopus citations

Abstract

Selective inhibition of specific genes can be accomplished using genetic suppressor elements (GSEs) that encode antisense RNA, dominant negative mutant proteins, or other regulatory products. GSEs may correspond to partial sequences of target genes, usually identified by trial and error. We have used bacteriophage lambda as a model system to test a concept that biologically active GSEs may be generated by random DNA fragmentation and Identified by expression selection. Fragments from eleven different regions of lambda genome, encoding specific peptides or antisense RNA sequences, rendered E. coli resistant to the phage. Analysis of these GSEs revealed some previously unknown functions of phage lambda, including suppression of the cellular lambda receptor by an 'accessory' gene of the phage. The random fragment selection strategy provides a general approach to the generation of efficient GSEs and elucidation of novel gene functions.

Original languageEnglish (US)
Pages (from-to)711-717
Number of pages7
JournalNucleic acids research
Volume20
Issue number4
DOIs
StatePublished - Feb 25 1992
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Genetics

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