The use of antinucleoside antibodies to probe the organization of chromosomes denatured by ultraviolet irradiation.
The use of antinucleoside antibodies to probe the organization of chromosomes denatured by ultraviolet irradiation.
复制标题
使用抗核苷抗体来探测因紫外线照射而变性的染色体的组织。
DOI：
10.1016/0014-4827(74)90614-4
复制
发表时间：
1974
影响因子：
3.7
通讯作者：
O. J. Miller
中科院分区：
文献类型：
--
作者：
R. Schreck;Bernard F. Erlanger;O. J. Miller
紫外线照射甲醇：乙酸固定的人类和小鼠中期染色体使它们能够结合特异性嘌呤或嘧啶碱基的抗体。由于这些抗体与单链DNA反应，但不与天然DNA反应，我们的结果表明，紫外线照射在染色体DNA中产生单链区域。使用间接免疫荧光技术来检测抗体结合，观察到抗体结合的高度特征性、非随机模式。腺苷（抗A）和胸苷（抗T）抗体产生相同的结合模式，在大多数方面与奎纳克林产生的染色体带型相匹配。然而，在染色体1，9和16上的组成性异染色质的旁着丝粒区域中观察到了另外的强荧光灶，这些区域已通过原位DNA-RNA杂交显示为富含AT的人类卫星DNA的位置。胞苷抗体也与9号染色体的同一区域结合。在小鼠染色体制备物中，抗A和抗T都在含有着丝粒异染色质的区域产生明亮的荧光，该区域已被证明是该物种富含AT的卫星DNA的位置。
Ultraviolet irradiation of methanol: acetic acid-fixed human and mouse metaphase chromosomes rendered them capable of binding antibodies specific for purine or pyrimidine bases. Since these antibodies react with single-stranded but not with native DNA, our results indicate that UV irradiation generated single-stranded regions in chromosomal DNA. Using an indirect immuno-fluorescence technique to detect antibody binding, highly characteristic, nonrandom patterns of antibody binding were observed. Antibodies to adenosine (anti-A) and thymidine (anti-T) produced identical patterns of binding which in most respects matched the chromosome banding patterns produced by quinacrine. However, additional foci of intense fluorescence were seen in the paracentromeric regions of constitutive heterochromatin on chromosomes 1, 9 and 16, regions which had been shown by in situ DNA-RNA hybridization to be the locations of AT-rich human satellite DNA. Antibodies to cytidine also bound to the same region of chromosome 9. In mouse chromosome preparations, both anti-A and anti-T produced bright fluorescence of the region containing centromeric heterochromatin, which had been shown to be the location of the AT-rich satellite DNA of this species.