Niwa Takashi, Kawamura Yoshiaki, Katagiri Yoshihiro, Ezaki Takayuki
Department of Microbiology, Regeneration and Advanced Medical Science, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu, 501-1194, Japan.
J Microbiol Methods. 2005 May;61(2):251-60. doi: 10.1016/j.mimet.2004.12.006. Epub 2005 Jan 11.
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.
比较了裂殖酶和无色肽酶对细菌DNA/RNA提取的裂解活性。在用裂殖酶处理后再用SDS处理时,观察到许多细菌菌株能被快速裂解。裂殖酶和无色肽酶对具有A1α化学型(如绿色气球菌)和A3α化学型(如表皮葡萄球菌)的细菌菌株的细胞壁肽聚糖结构均表现出高裂解活性。对于具有A1γ化学型的菌株(如枯草芽孢杆菌),裂殖酶的裂解活性高于无色肽酶。裂殖酶的活性不会因NaCl浓度增加而受到不利影响。裂殖酶裂解物成功用于DNA和RNA的快速提取,而无色肽酶裂解物会降解RNA。获得的DNA和RNA成功用于16S rRNA扩增和实时RT-PCR检测。结论是裂殖酶可用于多种革兰氏阳性细菌的快速裂解,并可用于DNA/RNA分离方案。