Photoactivatable antisense DNA: suppression of ampicillin resistance in normally resistant Escherichia coli.
Photoactivatable antisense DNA: suppression of ampicillin resistance in normally resistant Escherichia coli.
复制标题
光激活反义 DNA：抑制正常耐药大肠杆菌中的氨苄青霉素耐药性。
DOI：
10.1089/ard.1991.1.117
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Wong,HH
中科院分区：
文献类型：
--
作者：
Gasparro,FP;Edelson,RL;O'Malley,ME;Ugent,SJ;Wong,HH
与β-内酰胺酶基因的一个片段互补并在特定位点含有青霉素单加合物的反义寡脱氧核糖核苷酸被检查其使正常耐药细菌对氨苄青霉素敏感的能力。用长波长紫外辐射（380-400 nm）照射寡核苷酸和peptides产生单加合物反义分子。用高效液相色谱法纯化微克量的光活化反义DNA，用含β-内酰胺酶基因的质粒转化大肠杆菌，在长波长紫外线（320-400 nm）的光活化作用下，检测一系列含有补骨脂素单加合物的寡核苷酸。用这种光活化形式的反义DNA（0.4 μM）处理的正常耐药细菌对氨苄青霉素特异性致敏。与不含可光活化单加合物部分的对照寡核苷酸相比，菌落形成的减少范围为31%至79%。以类似方式处理的细菌，但在四环素而不是氨苄青霉素的存在下，没有受到影响。β-半乳糖苷酶基因与β-内酰胺酶基因位于同一质粒上，酶活性不受影响。
Antisense oligodeoxyribonucleotides complementary to a segment of the β-lactamase gene and containing psoralen monoadducts at specific sites were examined for their ability to make normally resistant bacteria sensitive to ampicillin. Irradiation of oligonucleotides and psoralens with long-wavelength ultraviolet radiation (380–400 nm) produced monoadducted antisense molecules. High-performance liquid chromatography was used to purify microgram quantities of photoactivatable antisense DNA.Escherichia colitransformed with a plasmid containing the gene for β-lactamase were used to test a series of oligonucleotides containing psoralen monoadducts after additional exposure to the photoactivating effects of long-wavelength ultraviolet radiation (320–400 nm). Normally resistant bacteria treated with this photoactivatable form of antisense DNA (0.4 μM) were specifically sensitized to ampicillin. The reduction in colony formation ranged from 31 to 79% in comparison to control oligonucleotides which did not contain photoactivatable monoadduct moieties. Bacteria treated in a similar manner but in the presence of tetracycline instead of ampicillin were not affected. The activity of β-galactosidase, whose gene is located on the same plasmid as β-lactamase, was not affected.