【摘要】 目的选取合适的靶点,探讨利用反义技术抑制结核杆菌的新方法。方法选取分枝杆菌ino1基因为靶基因,设计并合成3段反义寡核苷酸,将不同浓度的反义寡核苷酸加入细菌的培养液中,观察ino1的mRNA表达情况,结核杆菌分枝硫醇的水平以及结核杆菌的生长情况。结果当反义寡核苷酸的浓度达到20μmol/L时,可以特异性地抑制ino1的表达,此浓度水平时ino1 mRNA的量和对照组的比值为0.67,40μmol/L时的比值则为0.45。分枝硫醇的合成以及分枝杆菌的生长也相应地受到抑制。反义寡核苷酸的浓度为20μmol/L时,分枝杆菌浓度的对数值下降1.3,40μmol/L时下降1.7。结论利用反义技术抑制分枝杆菌是完全可行的,并且ino1可以作为反义寡核苷酸的靶基因。更多还原

【Abstract】 Objective To evaluate the use of antisense phosphorothioate-modified oligonucleotides(PS-ODNs) complementary to Mycobacterium tuberculosis(M.tuberculosis) as a possible antimycobacterial strategy.Methods M.tuberculosis inositol-1-phosphate synthetase(ino1) was chosen as the target of antisense oligonucleotides.Three antisense oligonucleotides were synthesized and added to the culture of M.tuberculosis to investigate the effect on the mRNA expression of ino1,level of mycothiol and the proliferation of M.tuberculosis.Results At the concentration of 20 μmol/L,the antisense oligonucleotides could inhibit the mRNA expression of ino1.The fold change of ino1 mRNA in M.tuberculosis treated with 20 μmol/L antisense oligonucleotides relative to M.tuberculosis incubated with medium alone was 0.67,and that of 40 μmol/L antisense oligonucleotides was 0.45.The level of mycothiol and the proliferation of M.tuberculosis were also inhibited.At concentration of 20 μmol/L,antisense oligonucleotides decreased growth by 1.3 log units,and that of 40 μmol/L was 1.7 log units.Conclusion M.tuberculosis ino1 is a target for the development of novel antibiotics and PS-ODN to ino1 is a promising antimycobaterial candidate.更多还原