摘要
【目的】确定鸡源鲍氏志贺菌的进化地位。【方法】选用6条随机引物（P1～P6）,采用随机扩增多态性DNA（RAPD）技术,对1株鸡源鲍氏志贺菌、2株鸡白痢沙门菌、2株鸡肠致病性大肠杆菌和2株痢疾志贺菌共7株菌的基因组DNA进行分析,同时对个别特殊序列进行克隆测序,并在此基础上分析了鸡源鲍氏志贺菌与其他15个相关菌株的进化关系。【结果】6条随机引物中有4条引物（P1,P2,P4和P6）能较好地在志贺菌、鸡白痢沙门氏菌和鸡致病性大肠杆菌3种菌中检测到多态性分子标记;这4条有效引物共扩增出了64个DNA片段,其中7个菌株共有的谱带有3条,多态性片段有61条,多态率达95.3%。引物P2可根据扩增图谱将志贺菌和大肠杆菌与沙门菌鉴别出来;用引物P6对上述7株细菌进行扩增,发现同种菌株间谱带特别相似,其中鸡鲍氏志贺菌与人痢疾志贺菌扩增条带的相同率达66.7%,与鸡肠致病性大肠杆菌扩增条带的相同率达44.4%,而鸡白痢沙门菌与志贺菌的扩增谱带相差甚远,可用于3种细菌的鉴别。对扩增片段进行的测序及进化分析结果显示,鸡源鲍氏志贺菌与人源志贺菌的进化距离最近。【结论】鸡源鲍氏志贺菌可能是人源志贺菌的一个变异株或新的亚种。
[Objective] The evolutionary position of chicken Shigella boydii was determined. [Method] The genetic diversity among strains of chicken Shigella boydii, S. Pullorum, Enteroinvasive E. coli and Shigella Dysenteriae was studied by random amplified polymorphic DNA （RAPD） with 6 random primers, and some special PCR products were cloned and sequenced, Further evolutionary relationships among chicken Shigella boydii and other related strains were examined based on sequences. [Result] Four efficient primers were selected from six 10 bp random primers. 64 RAPD bands were identified and 3 of them were common in all three species tested and 61 of them were polymorphic between any two strains. The percentage of polymorphism was 95.3 %. The primer P2 could be used to differentiate chicken Shigella boydii and S. Pullo-rum from four bacterial strains. The result showed that the common rate of the amplified bands between Shigella boydii and Shigella dysenteriae was 66.7 % ,Shigella boydii and Enteroinvasive E. coli was 44.4% ,but the amplified bands produced by Shigella and S. Pullorum was much different from each other when primer P6 was used,so the primer p6 could be used to differentiate the three species. The results for sequenced PCR products and examined evolutionary relationships showed that the chicken Shigella boydii may be a new species or subspecies from human. [Conclusion] The chicken Shigella boydii may be a new species or subspecies from human.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2009年第10期12-18,共7页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家自然科学基金项目(30671555)
