用双示踪法分析质粒DNA合成——利福平与氯霉素对大肠杆菌K-12中pSC101和ColEl-amp复制的不同影响
Analysis of plasmid DNA synthesis by double tracer method--differential effects of rifampicin and chloramphenicol on the replication of pSC101 and ColEl-amp in Escherichia coli K-12.
作者信息
Hasunuma K, Furukawa K
出版信息
Radioisotopes. 1983 Dec;32(12):609-16. doi: 10.3769/radioisotopes.32.12_609.
An Escherichia coli strain, CR34 , harboring both pSC101 and ColEl -amp plasmids was exposed to media containing rifampicin (100 micrograms/ml) and/or chloramphenicol (180 micrograms/ml) and the cells were labeled for 20 min with 3H-thymine at 3, 25 and 50 min after exposure to drug(s). The plasmid DNA synthesis was assayed by DNA-DNA hybridization with 14C-labeled pSC 134 DNA as internal marker. In the presence of rifampicin, the replication of pSC 101 was from 57 to 104% that in its absence, and that of ColEl -amp was from 17 to 26%. The DNA replication of pSC 101 after addition of chloramphenicol was reduced to 35 to 75%, and that of ColEl -amp was reduced to 39% and then restored to 92%. This restoration was not observed in the presence of rifampicin.
一种同时携带pSC101和ColEl - amp质粒的大肠杆菌菌株CR34，被置于含有利福平（100微克/毫升）和/或氯霉素（180微克/毫升）的培养基中，并在接触药物后3、25和50分钟用³H - 胸腺嘧啶对细胞进行20分钟的标记。通过以¹⁴C标记的pSC 134 DNA作为内部标记进行DNA - DNA杂交来检测质粒DNA的合成。在有利福平存在的情况下，pSC 101的复制是其不存在时的57%至104%，ColEl - amp的复制是其不存在时的17%至26%。添加氯霉素后，pSC 101的DNA复制降至35%至75%，ColEl - amp的DNA复制降至39%，然后恢复到92%。在有利福平存在的情况下未观察到这种恢复情况。
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