Open AccessAssessment of the utilization of the antisense RNA strategy to identify essential genes in heterologous bacteria
Author(s) -
Wang Bing,
Kuramitsu Howard K
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00128-9
Subject(s) - biology , escherichia coli , gene , cloning (programming) , streptococcus mutans , bacteria , antisense rna , genetics , heterologous expression , heterologous , plasmid , rna , dna , microbiology and biotechnology , recombinant dna , computer science , programming language
We employed an antisense RNA approach to identify essential genes common in both Gram‐positive and Gram‐negative bacteria by cloning a random library of Streptococcus mutans chromosomal DNA into an expression vector and transforming Escherichia coli . Twelve out of 27 E. coli transformants with growth defective phenotypes contained individual structural genes of S. mutans in the antisense orientation relative to the E. coli promoter. Thirty‐three percent of these transformants (4/12) corresponded to the genes ( gyrA , ileS , rplE and yihA orthologs) which are essential for bacterial viability.