嗜水气单胞菌的分离鉴定及其Hfq功能的研究-轻工技术与工程专业论文.docxVIP

同表型测定发现，在对数生长期内，Ahfq突变株的生长相对于野生型有所 同表型测定发现，在对数生长期内，Ahfq突变株的生长相对于野生型有所 下降；在进入稳定期后，Ahfq突变株的生长达到与野生型相当的水平，并 在后期略微有所提高。Hfq对游动性具有正调控作用，对泳动性和溶血活性 调控不明显；当Hfq表达受损时，其胞外蛋白酶产量显著提高，即Hfq对 嗜水气单胞菌胞外蛋白酶的产量具有明显的抑制作用；此外，hfq基因缺失 后，嗜水气单胞菌在30％蔗糖和3％NaCl条件下的生长受到显著抑制：在 不同SDS和乙醇浓度下进行培养时，Ahfq突变株的生长亦受到一定程度的 影响。以斑马鱼为模型进行了嗜水气单胞菌感染实验，结果发现分离菌株具 有明显的毒力，当Hfq表达受损时，嗜水气单胞菌对斑马鱼的毒力显著下降。 关键词：嗜水气单胞菌，Hfq，突变株，毒力 万方数据 Isolation Isolation and Identification of Aeromonas hydrophila and Function Analasis of Hfq on Its Bioprocesses ABSTRACT Aeromonas hydrophila is a major pathogen causing the bacterial disease and brings about huge damage and economic losses in aquaculture all over the world． Meanwhile，this bacterial，a zoonotic pathogen，can also infect human and is a threat to human health．Thus，to study the virulence regulation of this bacterial will do some favor to the control of its resulting disease．Hfq．the sRNA chaperon and the global post—transcriptional regulator,is widespreaded in bacteria and involved in various bioprocesses and virulence regulation in kinds of pathogensa by promoting the interaction of different sRNAs and their target mRNAs．In this study,a pathogenic strain，isolated from the diseased grouper in the fish farm in Yantai，Shandong Province，was characterized to be A． hydrophila by physiological，biochemical，and molecular methods．Then the hfq gene in this strain was cloned and the彳蜘mutant strain was constructed by markerless gene deletion strategies．Furthermore，the regulation of Hfq on the physiological functions and virulence in A．hydrophila was figured out．This will broaden the knowledgment about the virulence regulation mechanism in A． hydrophila and also lay some theorial foundation for scientific prevention of this pathogen．The results were just as follows： (1)A dominant strain was isolated from the pancreatic tissue of the diseased grouper．The strain，which was Gram negative short—rod bacterial，Can form D hemolysis circle，utilize glucose to generate acid and gas，reduce the nitrate， dec