CAJ | 학술논문

应用分子生物学技术，开展鼠疫菌调控子蛋白 OxyR 对 rovAR 的转录调控机制研究。提取鼠疫菌野生株（WT）和 oxyR 突变株（ΔoxyR）的总 RNA，采用引物延伸试验研究 rovAR 的转录起始位点，并根据产物的丰度判断 OxyR 对 rovAR 的调控关系。构建克隆有 rovAR 上游启动子区的 lacZ 重组质粒，将重组质粒转入 WT 和ΔoxyR 中，通过测定β-半乳糖苷酶活性来比较2株重组菌中的 rovAR 启动子活性，进一步验证 OxyR 对 rovAR 的调控关系。进而利用大肠埃希菌 OxyR 识别基序，预测 OxyR 对 rovAR 启动子区的结合位点。发现 rovAR 的2个转录起始位点均受 OxyR 的激活，且 rovAR 启动子区具有可能的 OxyR结合位点。推测 OxyR 能直接结合到 rovAR 启动子区而激活其转录表达。
응용분자생물학기술，개전서역균조공자단백 OxyR 대 rovAR 적전록조공궤제연구。제취서역균야생주（WT）화 oxyR 돌변주（ΔoxyR）적총 RNA，채용인물연신시험연구 rovAR 적전록기시위점，병근거산물적봉도판단 OxyR 대 rovAR 적조공관계。구건극륭유 rovAR 상유계동자구적 lacZ 중조질립，장중조질립전입 WT 화ΔoxyR 중，통과측정β-반유당감매활성래비교2주중조균중적 rovAR 계동자활성，진일보험증 OxyR 대 rovAR 적조공관계。진이이용대장애희균 OxyR 식별기서，예측 OxyR 대 rovAR 계동자구적결합위점。발현 rovAR 적2개전록기시위점균수 OxyR 적격활，차 rovAR 계동자구구유가능적 OxyR결합위점。추측 OxyR 능직접결합도 rovAR 계동자구이격활기전록표체。
To stuy the transcriptional regulation mechanism of rovA by OxyR inYersinia pestis ,total bacte-rial RNAs were extracted from the wide-type (WT)strain and the oxyR null mutant (ΔoxyR).Primer ex-tension assay was employed to detect the promoter activity (the amount of primer extension product)of ro-vA in WT and that in ΔoxyR.The rovA promoter-proximal region was cloned into the pRW50 containing a promoterless lacZ gene.The recombinant LacZ reporter plasmid was transformed into WT and ΔoxyR, respectively,to measure the promoter activity (theβ-Galactosidase activity)of rovA in WT and ΔoxyR by using theβ-Galactosidase Enzyme Assay System.And then the E.coli OxyR consensus was carried out to predict the OxyR binding site to rovA promoter region.All the two transcription starts of rovA were acti-vated by OxyR through directly bind to the promoter sequences.OxyR may directly activate the rovA tran-scription inYersinia pestis .