RRC ID 1144
Author Niwa T, Kawamura Y, Katagiri Y, Ezaki T.
Title Lytic enzyme, labiase for a broad range of Gram-positive bacteria and its application to analyze functional DNA/RNA.
Journal J. Microbiol. Methods
Abstract The lytic activity of labiase and achromopeptidase for bacterial DNA/RNA extraction were compared. Rapid lysis of many bacterial strains was observed with labiase followed by SDS treatment. Both labiase and achromopeptidase showed high lytic activity against bacterial strains with the A1alpha chemotype (e.g., Aerococcus viridans) and the A3alpha chemotype (e.g., Staphylococcus epidermidis) for cell wall peptidoglycan structures. The lytic activity of labiase was higher than that of achromopeptidase against strains with the A1gamma chemotype (e.g., Bacillus subtilis). The activity of labiase was not detrimentally affected with increasing NaCl concentration. Labiase lysates were successfully used for rapid extraction of DNA and RNA, whereas achromopeptidase lysates degraded RNA. The DNA and RNA obtained were successfully used for 16S rRNA amplification and real-time RT-PCR detection. It is concluded that labiase is useful for rapid lysis of a wide variety of Gram-positive bacteria and can be used for DNA/RNA isolation protocols.
Volume 61(2)
Pages 251-60
Published 2005-5
DOI 10.1016/j.mimet.2004.12.006
PII S0167-7012(04)00361-6
PMID 15722152
MeSH Bacterial Outer Membrane Proteins / metabolism* Bacteriolysis / drug effects Bacteriolysis / physiology DNA, Bacterial / genetics DNA, Bacterial / isolation & purification* Endopeptidases / metabolism* Gram-Positive Bacteria / genetics Gram-Positive Bacteria / metabolism* RNA, Bacterial / genetics RNA, Bacterial / isolation & purification* Reverse Transcriptase Polymerase Chain Reaction Serine Endopeptidases / metabolism
IF 1.701
Times Cited 23
WOS Category BIOCHEMICAL RESEARCH METHODS MICROBIOLOGY
Resource
Pathogenic microorganisms A. viridans? S. epidermidis? B. subtilis?