Assessment of the utilization of the antisense RNA strategy to identify essential genes in heterologous bacteria.
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.
Wang
Bing
B
Department of Oral Biology, State University of New York at Buffalo, Buffalo, NY 14212, USA.
Kuramitsu
Howard K
HK
D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics; D020043:Genes, Essential / Q000235:genetics; D006090:Gram-Negative Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006094:Gram-Positive Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D008957:Models, Genetic; D009154:Mutation; D016372:RNA, Antisense / Q000235:genetics* / Q000502:physiology; D013295:Streptococcus mutans / Q000235:genetics / Q000302:isolation & purification
D016372:RNA, Antisense