伴放线放线杆菌粗糙型与光滑型菌株生物学特性的初步研究

【作者】 张晶；

【导师】 钟德钰；

【作者基本信息】 青岛大学 ， 口腔临床医学， 2010， 硕士

【摘要】 目的：1.研究伴放线放线杆菌同一菌株粗糙型和光滑型的生长曲线趋势。2.研究伴放线放线杆菌同一菌株粗糙型与光滑型菌株的菌体蛋白表达差异。方法：1.采用液体培养基TSB培养伴放线放线杆菌临床分离株D7S及其光滑型变异菌株D7SS,用紫外分光光度计检测细菌密度,连续观测30h,分别绘制生长曲线。2.将伴放线放线杆菌临床菌株D7S及其光滑型菌株D7SS细菌全细胞进行聚丙烯酰胺凝胶(SDS-PAGE)电泳,考马斯亮蓝染色后比较两种表型的细菌蛋白表达的差异。采用免疫印迹法(Western Blot)观察两种表型细菌蛋白对单克隆抗体TEPC-15的反应情况。结果：1、D7SS菌株比D7S菌株更早进入对数期和衰亡期；2、D7SS菌株和D7S菌株细菌全细胞蛋白电泳考马斯亮蓝染色时蛋白带未见明显差异；3、免疫印迹分析D7S菌株在分子量接近lOKu处存在对TEPC-15特异性反应的磷酸胆碱,而在D7SS菌株中未发现。结论：伴放线放线杆菌D7S和其光滑型菌株D7SS在液体培养基TSB中的生长曲线不同；D7S和D7SS聚丙烯酰胺凝胶(SDS-PAGE)电泳后对TEPC-15抗体的反应性不同。更多还原

【Abstract】 Objective:1. To draw growth curve of two different phenotypes of Aggregatibacter actinomycetemcomitans(Aa) from the same strain.2. To detect possible differences in the two bacterial protein expression.Methods:1. Aggregatibacter actinomycetemcomitans clinical isolates D7S and its smooth variant strain D7SS were cultured in the liquid culture medium TSB, and draw the growth curve after detecting the two bacterial strains’density every 2 hours during 30 hours using UV Spectrophotometer. 2. Compare the bacterial proteins by Coomassie brilliant blue after Polyacry lamide gel electrophoresis. And observe reaction to monoclonal antibody TEPC-15 by Western blot analysis.Results:1.Aa D7SS strain entered into both logarithmic phase and decline phase earlier than D7S strain.2 There were no significant differences between their protein bands stained by Coomassie brilliant blue after whole-cell protein electrophoresis; 3. The analysis of Western Blot showed that Phosphorylcholine were found in D7S whose molecular weight was about 10Ku, but not in D7SS.Conclusion:The research revealed that there were significant differences in the growth curve and reactivity with TEPC-15 antibody between Aggregatibacter actinomycetemcomitans D7S and its smooth variant strain D7SS.更多还原