温度和酸度对田鼠型鼠疫耶尔森菌psa位点的转录调控研究
发布时间：2019-04-02 10:31
【摘要】：目的研究温度和酸度对田鼠型鼠疫耶尔森菌psa位点的转录调控作用。方法首先设计相关跨基因引物,用RT-PCR方法验证psa位点的相关操纵子结构。扩增操纵子首基因上游启动子区500 nt序列,并克隆入pRW50质粒β-半乳糖苷酶基因的上游,将重组质粒转入鼠疫菌中,通过测定β-半乳糖苷酶活性差异来判断不同温度(26和37℃)和酸度(pH 6.0和pH 8.0)对psaE和psaA的调控关系。最后提取鼠疫菌在上述不同温度和酸度条件下的总RNA,采用引物延伸实验进一步明确温度和酸度对psaA的调控关系。结果与结论 RT-PCR结果证实了田鼠型鼠疫菌的psa位点由操纵子psaEF和psaABC构成;半乳糖苷酶和引物延伸实验结果显示在37℃、pH 6.0培养条件下psaABC的转录水平最高,而psaEF的转录水平无明显变化,提示psaA的表达水平在37℃酸性条件下表达量最高,psaE则不受温度和酸度的调控。
[Abstract]:Objective To study the effect of temperature and acidity on the transcription and control of the psa site in the microvole-type Yersinia pestis. Methods The relative operon structure of the psa site was verified by RT-PCR. the sequence of the promoter region of the upstream promoter region of the first gene of the operon is amplified and cloned into the upstream of the pRW50 plasmid pRW50 plasmid, The regulation and control of psaE and psaA were determined by the determination of the difference of the activity of the yeast-galactooligosaccharides (pH 6.0 and pH 8.0) at different temperatures (26 and 37.degree. C.) and the acidity (pH 6.0 and pH 8.0). Finally, the total RNA of Yersinia pestis under different temperature and acidity conditions was extracted, and the control relationship of temperature and acidity to psa A was further determined by the primer extension experiment. The results and conclusions RT-PCR confirmed that the psa site of the microvole-type Yersinia pestis was composed of the operon psaEF and psaABC. The results showed that the transcription level of psaABC was the highest at 37 鈩，
本文编号：2452485
[Abstract]:Objective To study the effect of temperature and acidity on the transcription and control of the psa site in the microvole-type Yersinia pestis. Methods The relative operon structure of the psa site was verified by RT-PCR. the sequence of the promoter region of the upstream promoter region of the first gene of the operon is amplified and cloned into the upstream of the pRW50 plasmid pRW50 plasmid, The regulation and control of psaE and psaA were determined by the determination of the difference of the activity of the yeast-galactooligosaccharides (pH 6.0 and pH 8.0) at different temperatures (26 and 37.degree. C.) and the acidity (pH 6.0 and pH 8.0). Finally, the total RNA of Yersinia pestis under different temperature and acidity conditions was extracted, and the control relationship of temperature and acidity to psa A was further determined by the primer extension experiment. The results and conclusions RT-PCR confirmed that the psa site of the microvole-type Yersinia pestis was composed of the operon psaEF and psaABC. The results showed that the transcription level of psaABC was the highest at 37 鈩，
本文编号：2452485
本文链接：https://www.wllwen.com/yixuelunwen/jichuyixue/2452485.html
