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在中国仓鼠卵巢细胞中使用单克隆抗体在极低溴脱氧尿苷替代水平下测量姐妹染色单体交换。

Measurement of sister chromatid exchanges at very low bromodeoxyuridine substitution levels using a monoclonal antibody in Chinese hamster ovary cells.

作者信息

Pinkel D, Thompson L H, Gray J W, Vanderlaan M

出版信息

Cancer Res. 1985 Nov;45(11 Pt 2):5795-8.

PMID:4053051
Abstract

A monoclonal antibody to bromodeoxyuridine (BrdUrd) incorporated into DNA allowed visualization of sister chromatid exchanges (SCE) when as little as 0.6% of the thymine in a single DNA strand has been substituted. Measurement of the SCE frequency as a function of BrdUrd substitution in a normal Chinese hamster ovary cell line showed a plateau of six SCEs per cell for substitution levels up to at least 20%. A clear elevation in frequencies was noted at 60% substitution. However, in the mutant line EM9, previously shown to have a highly elevated frequency of SCE, the level of exchanges declined continuously as the percentage of BrdUrd substitution decreased. At 0.6% substitution, the frequency of SCE was still 4-fold higher than that of the parental cells. The antibody procedure described here should be useful in evaluating the extent to which SCEs induced by mutagenic agents result from interactions between the DNA damage caused by the agent and the BrdUrd routinely used for measuring SCE.

摘要

一种针对掺入DNA中的溴脱氧尿苷(BrdUrd)的单克隆抗体,当单链DNA中胸腺嘧啶的取代量低至0.6%时,可使姐妹染色单体交换(SCE)可视化。在一个正常的中国仓鼠卵巢细胞系中,测量SCE频率作为BrdUrd取代量的函数,结果显示,对于高达至少20%的取代水平,每个细胞有6次SCE的平台期。在60%取代时,频率明显升高。然而,在先前已显示SCE频率高度升高的突变系EM9中,随着BrdUrd取代百分比的降低,交换水平持续下降。在0.6%取代时,SCE频率仍比亲代细胞高4倍。本文所述的抗体方法应有助于评估诱变剂诱导的SCE在多大程度上是由该试剂引起的DNA损伤与常规用于测量SCE的BrdUrd之间的相互作用导致的。

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