论文部分内容阅读
作者通过机械破裂和化学提取制备卡介苗(BCG)基因组文库,再用酶将DNA消化成30～50kb片段,与质粒pYUB178整合后用电穿孔术插入耻垢分枝杆菌mc~2155中,用卡那霉素琼脂选出约250个转化体作为带有BCG基因组的重组耻垢分枝杆菌库(J3R)。同时取未整合BCG基因的pYUB178转化体作对照菌。作者首先将约10~6菌落形成单位(cfu)J3R和对照菌各0.2ml分别静脉内感染无特殊病原体的C57BL/6小鼠,在14天内间隔不同时间取脾和肺组织匀浆接种于琼脂培养基进行菌落计数。结果显示,J3R感染小鼠的脾内活菌数随时间增长而逐渐减少,但略比对照菌感染小鼠的脾内活菌数多。
The authors prepared a Bacillus Calmette-Guerin (BCG) genomic library by mechanical disruption and chemical extraction, digested the DNA into 30-50 kb fragments with an enzyme, integrated the plasmid pYUB178 and electroporated into M. smegmatis mc-2155, Approximately 250 transformants were selected for mycotoxin agar as a recombinant M. smegmatis pool (J3R) with the BCG genome. Meanwhile, pYUB178 transformant without BCG gene was used as a control. At first, C57BL / 6 mice without specific pathogens were intravenously infected with about 10 ~ 6 colony forming units (cfu) J3R and 0.2ml each of the control bacteria, and spleen and lung tissue homogenates were inoculated on agar at different time intervals within 14 days Culture medium for colony counting. The results showed that the number of viable cells in the spleen of J3R-infected mice gradually decreased over time, but slightly more than the viable cells in the spleen of mice infected with control bacteria.