We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
禽多杀性巴氏杆菌C48-1株胞外分泌蛋白的DNase活性分析.
- Authors
陈耀华; 董何凡; 李 琦; 牛俊辉; 钱 满; 王晓利; 全映颖; 刘桂坤; 雷显前; 程相朝; 廖成水
- Abstract
To investigate the DNase activity of the extracellular secreted proteins from Pasteurella multocida (Pm) and the factors that affect the DNase activity. In this study, the degradation of λDNA by the extracellular secreted proteins from avian Pm C48-1 strain was determined by agar culture assay. The extracellular secreted proteins of avian Pm C48-1 strain was collected by low-temperature dialysis after 5 hours of culture and the concentrations of the proteins were determined by BCA kit. The DNase activity level of the extracellular secreted proteins from Pm was detected by the Kunitz method. The result showed that the extracellular secreted proteins of Pm exhibited DNase activity that degrades λDNA. The concentration of the proteins was 1.32μg/mL and the activity level of degrading λDNA was 6.07×106 Kunitz Unit/mg. The ability of the degradation of different DNA substrates and the effect of acid/base, temperature, and different concentrations of metal cations on the DNase activity of the extracellular secreted proteins were analyzed by agarose gel electrophoresis. The results showed that the DNase activity of the extracellular secreted proteins had a broad spectrum that can degrade λDNA, bacterial genomic DNA, plasmid DNA, and PCR-amplified DNA fragments. The detection results of the factors affecting DNase activity showed that the optimal reaction temperature for degrading λDNA was 30°C. The DNase activity was better at pH7.0, pH8.0, and pH9.0, however, there was no DNase activity under acidic and strongly alkaline conditions. The results of agarose gel electrophoresis also showed that Na+, K+, and Ba2+ in the concentrations of 0.01mmol/L, 0.1mmol/L, 1mmol/L, and 5mmol/L had no significant effect on the DNase activity; Ca2 +, Mg2 +, and Mn2 + increased the DNase activity of extracellular secreted proteins; Co2 + in the concentrations of 1mmol/L and 5mmol/L and Cu2+ in the concentrations of 0.01mmol/L and 0.1mmol/L showed inhibitory effects, while Cu2+ in the concentrations of 1mmol/L and 5mmol/L had no significant effect on the DNase activity. The present study, for the first time, confirmed the DNase activity of the extracellular secreted proteins of avian Pm C48-1 strain, which had a broad spectrum, and identified the factors affecting the DNase activity of the extracellular secreted proteins. This study provided a reference for further exploring the role of the extracellular DNase in the immune interaction between Pm infection and host.
- Subjects
BACTERIAL DNA; PASTEURELLA multocida; GEL electrophoresis; AGAROSE; PROTEINS
- Publication
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao, 2022, Vol 44, Issue 9, p984
- ISSN
1008-0589
- Publication type
Article
- DOI
10.3969/j.issn.1008-0589.202111008