Characterization of antibodies to dihydrothymine, a radiolysis product of DNA.
Characterization of antibodies to dihydrothymine, a radiolysis product of DNA.
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二氢胸腺嘧啶（DNA 的放射分解产物）抗体的表征。
DOI：
10.1021/bi00436a039
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发表时间：
1989
期刊：
影响因子：
2.9
通讯作者：
Wallace,SS
中科院分区：
文献类型：
--
作者：
Hubbard,K;Ide,H;Erlanger,BF;Wallace,SS
凯伦·哈伯德，1，8井出浩，1，11伯纳德·F。埃尔朗格，1和苏珊S。Wallace**-1，11微生物学和免疫学系，纽约医学院，瓦尔哈拉，纽约10595，和微生物学系，哥伦比亚大学内科医生和外科医生学院，纽约，纽约10032收到1988年11月17日;修订稿收到1989年1月27日摘要：用碳二亚胺偶联牛血清白蛋白（BSA）的二氢胸苷单磷酸免疫家兔，获得了抗二氢胸苷的抗体。通过使用ELISA测定，发现产生的抗体对二氢胸腺嘧啶具有特异性。半抗原抑制研究表明，二氢胸苷一磷酸作为抑制剂比胸苷一磷酸有效3个数量级，比胸苷乙二醇一磷酸有效4个数量级。对于含有二氢胸腺嘧啶的DNA，在20 fmol的二氢胸腺嘧啶下观察到抗体反应性，其约为每10000个碱基0.1个二氢胸腺嘧啶。因此，该测定非常灵敏。该抗体与含有二氢胸腺嘧啶的变性DNA反应，但不与含有该病变的天然DNA反应。该抗体用于测量在野生型大肠杆菌或突变体中体内掺入的二氢胸苷，这些突变体在从DNA中去除二氢胸苷或从头合成胸苷酸的能力方面存在缺陷。最后，用二氢胸腺嘧啶抗体定量分析了N_2下X射线照射DNA中二氢胸腺嘧啶的形成。生产二氢胸腺嘧啶在辐照的DNA与减少由X-射线产生的物种的水平，和二氢胸腺嘧啶优先产生在辐照的单链或变性的DNA相比，辐照的双链体DNA。
Karen Hubbard, 1, 8 Hiroshi Ide, 1, 11 Bernard F. Erlanger, 1 and Susan S. Wallace**-1, 11 Department of Microbiology and Immunology, New York Medical College, Valhalla, New York 10595, andDepartment of Microbiology, Columbia UniversityCollege of Physicians and Surgeons, New York, New York 10032 Received November 17, 1988; Revised Manuscript Received January 27, 1989 abstract: Antibodies to dihydrothymine were elicited by immunizing rabbits with dihydrothymidine monophosphate conjugated by carbodiimide to BSA. By use of an ELISA assay, the antibodies produced were found to be specific for dihydrothymine. Hapten inhibition studies showed that dihydrothymidine monophosphate was 3 orders of magnitude more effective as an inhibitor than thymidine monophosphate and 4 orders of magnitude more effective than thymidine glycol monophosphate. With DNA containing dihydrothymine, antibody reactivity was observed at 20 fmol of dihydrothymine, which is approximately 0.1 dihydrothymine per 10000 bases. Thus, the assay is very sensitive. The antibody reacted with denatured DNA containing dihydrothymine but not with native DNA containing this lesion. The antibody was used for measurement of in vivo incorporation of dihydrothymidine in wild-type Escherichia coli or mutants defective in their ability to remove dihydrothymine from DNA or in the de novo synthesis of thymidylate. Lastly, antibodiesto dihydrothymine were used to quantitate the formation of dihydrothymine in DNA X-irradiated under N2. Production of dihydrothymine in irradiated DNA correlated with the level of reducing species produced by X-rays, and dihydrothymine was produced preferentially in irradiated single-stranded or denatured DNA as compared to irradiated duplex DNA.