CAJ | 학술논문

目的：：用原核系统表达结核分枝杆菌Rv3425蛋白并纯化，评价该重组蛋白在结核病血清学诊断方面的价值。方法：以结核分枝杆菌H37Rv株基因组为模板，PCR扩增得到Rv3425基因序列，克隆至表达载体pET-28a中，转入大肠杆菌BL21（DE3）进行诱导、表达后纯化，用Western印迹和ELISA法进行抗原性初步评价。结果：在原核系统内经IPTG诱导表达后，Rv3425蛋白主要以包涵体形式存在，经复性和镍柱层析纯化后，纯度达95%以上；Western印迹和ELISA结果证明重组Rv3425具有较强的抗原活性；用纯化的Rv3425蛋白做抗原，临床诊断结核病人血清，阳性率达50%。结论：高纯度的Rv3425蛋白在结核病诊断方面具有很高的应用价值，可作为结核病诊断的备选抗原。
목적：：용원핵계통표체결핵분지간균Rv3425단백병순화，평개해중조단백재결핵병혈청학진단방면적개치。방법：이결핵분지간균H37Rv주기인조위모판，PCR확증득도Rv3425기인서렬，극륭지표체재체pET-28a중，전입대장간균BL21（DE3）진행유도、표체후순화，용Western인적화ELISA법진행항원성초보평개。결과：재원핵계통내경IPTG유도표체후，Rv3425단백주요이포함체형식존재，경복성화얼주층석순화후，순도체95%이상；Western인적화ELISA결과증명중조Rv3425구유교강적항원활성；용순화적Rv3425단백주항원，림상진단결핵병인혈청，양성솔체50%。결론：고순도적Rv3425단백재결핵병진단방면구유흔고적응용개치，가작위결핵병진단적비선항원。
Objective: To obtain protein Rv3425 of Mycobacterium tuberculosis and to evaluate its potential value for tuberculosis serological diagnosis. Methods: Rv3425 gene was obtained by PCR from genome of M.tuberculosis H37Rv and was cloned into pET-28a vector. After plasmid transformation and IPTG induction, the recombinant protein was expressed. The antigenicity of the purified product were examined by Western blotting and ELISA. Re-sults: Recombinant protein Rv3425 was expressed in form of inclusion body and the purity reached 95% after re-nature and purification. The strong antigenic ability of Rv3425 was detected by Western blotting and ELISA test, the positivity was about 50% with tuberculosis serological diagnosis. Conclusion: Recombinant Rv3425 protein had an important value in diagnosis of tuberculosis, and it might be diagnostic antigen candidate of tuberculosis.