中国兽医科学2024，Vol.54Issue(1)：41-47,7.DOI:10.16656/j.issn.1673-4696.2024.0010
绵羊肺炎支原体荚膜多糖的提取纯化及反应原性分析
Extraction,purification and reactivity analysis of capsular polysaccharide of Mycoplasma ovipneumoniae
摘要
Abstract
This study aimed to extract the capsular polysaccharides from GH3-3 strain of Mycoplasma ovipneumoniae and verify its reactivity.Crude capsular polysaccharide was obtained by absolute ethanol precipitation method,then nucleic acid and protein were removed by phenol extraction and en-zyme treatment to obtain purified capsular polysaccharide.Immunofluorescence test was used to prove the reactivity of the capsular polysaccharide,and Congo red test was carried out to verify the triple helix structure of the capsular polysaccharides.12.24 mg of crude capsular polysaccharides derived from Mo GH3-3 was initially extracted from the supernatant of 1 L medium,and 9.8 mg of refined capsular polysaccharide was obtained after purification.The purified yield was about 80％.The results of im-munofluorescence experiment showed that Mo GH3-3 capsule polysaccharides had good reactivity.The Con-go red assay confirmed that the capsular polysaccharides of Mo has a triple helical structure.The high purity capsular polysaccharides of Mo GH3-3 strain were obtained using a combination of technical methods,comprising alcohol precipitation,phenol extract ion,enzyme hydrolysis,ultrafiltration and concentration.This extraction and purification assay for Mo capsular polysaccharides is not only sim-ple but also cost-effective,making it is suitable for application in production practice.The capsular polysaccharides exhibit good reactivity and have a triple helix structure,indicating that its biolo-gical activity is well-preserved.This work provides a reference method for the preparation of polysac-charide for the research and development of relevant diagnostic reagents,vaccines and adjuvants.关键词
绵羊肺炎支原体/荚膜多糖/三螺旋结构/反应原性Key words
Mycoplasma ovipneumoniae/capsular polysaccharide/triple helix structure/reactivity分类
农业科技引用本文复制引用
田彤彤,葛家振,李倩倩,高鹏程,吴晓妮,宋国栋,刘一健,郑福英,储岳峰..绵羊肺炎支原体荚膜多糖的提取纯化及反应原性分析[J].中国兽医科学,2024,54(1):41-47,7.基金项目
国家重点研发计划项目(2022YFD1800704,2022YFD1302101) （2022YFD1800704,2022YFD1302101）
甘肃省科技重大专项(22ZD6NA012) （22ZD6NA012）
中国农业科学院科技创新工程项目(CAAS-ASTIP-2021-LVRI) （CAAS-ASTIP-2021-LVRI）
