Abstract
A high throughput method for screening cDNA libraries has been developed to identify putative antimicrobial peptides (AMPs). It is based on a rapid dye inclusion assay for assessing antagonism of bacterial viability. Colonies are grown on a membrane on a permissive medium until full colony size is reached. The membrane, supporting the array of colonies, is transferred onto an inductive medium containing a vital dye. Upon expression of any antagonizing peptides, the cell membrane becomes compromised allowing dye infusion to permit visual identification of deleterious peptides. Our approach was validated by screening a synthetic oligonucleotide library expressed in Escherichia coli. A random oligonucleotide library, containing inserts of up to 75 nucleotides in length was constructed and expressed in E. coli. From a potential pool of 100000 peptides, in a single round of screening, three were found to be antimicrobial: L1, L3, and L8. Peptide L1 was shown to have a concentration-dependent bactericidal effect against Gram-negative E. coli and moderate biostatic activity against the Gram-positive bacteria Listeria monocytogenes. L8 was found to have bacteriostatic, and possibly bactericidal effect against E. coli, Pseudomonas aerug...Continue Reading
References
Mar 1, 1991·The Biochemical Journal·C L Ho, L L Hwang
Jul 16, 1981·Nature·H SteinerH G Boman
Aug 2, 1996·Journal of Medicinal Chemistry·M M JavadpourM L McLaughlin
Aug 1, 1997·Current Opinion in Structural Biology·P A Nygren, M Uhlén
Jan 29, 2000·European Journal of Biochemistry·J GorayaJ M Conlon
Jun 27, 2000·Bioinformatics·L J McGuffinD T Jones
Sep 2, 2000·Biophysical Journal·L Silvestro, P H Axelsen
Dec 12, 2001·Biochimica Et Biophysica Acta·K KonnoT Nakajima
Aug 17, 2002·Peptides·A L A RuissenA V Nieuw Amerongen
Sep 20, 2002·Journal of Medicinal Chemistry·Marina GobboRaniero Rocchi
Mar 5, 2003·Pharmacological Reviews·Michael R Yeaman, Nannette Y Yount
Jun 14, 2003·Journal of Virology·James S GibbsJonathan W Yewdell
Dec 19, 2003·Nucleic Acids Research·Zhe Wang, Guangshun Wang
May 30, 2006·Protein Science : a Publication of the Protein Society·Iddo FriedbergAdam Godzik
Aug 1, 2006·Journal of Peptide Science : an Official Publication of the European Peptide Society·Qiuhong XieSachiko Machida
Sep 8, 2006·The Biochemical Journal·Sónia Troeira HenriquesMiguel A R B Castanho
Mar 8, 2007·Bioinformatics·Christopher D FjellArtem Cherkasov
Nov 27, 2007·Nature Reviews. Molecular Cell Biology·David LeeChristine Orengo
Citations
Mar 23, 2011·Annals of Clinical Microbiology and Antimicrobials·Satoshi UenoYusuke Kato
Dec 22, 2010·Gut Pathogens·Filip Van Immerseel
Feb 5, 2010·International Journal of Antimicrobial Agents·Evelin LoitIllimar Altosaar
Dec 2, 2009·Journal of Peptide Science : an Official Publication of the European Peptide Society·Xiaojing ShenCui Hu
Jun 16, 2021·Toxicon : Official Journal of the International Society on Toxinology·Yi-Chuan MengKen Tan