Musarrat J, Ahmad M
Interdisciplinary Biotechnology Unit, Faculty of Life Sciences, Aligarh, India.
Biochem Int. 1991 Sep;25(2):249-59.
S1 nuclease hydrolysis, benzoylated naphthoylated DEAE cellulose (BND-cellulose) chromatography as well as certain immunological and genetic techniques have been used to evaluate the effect of mild alkali (pH10.0) on the DNA molecule. Native calf thymus DNA after exposure to alkaline pH10.0 when subjected to S1 nuclease hydrolysis released significant amount of acid soluble nucleotides as compared to the untreated control. With pBR322 DNA, the population of linear DNA species increased on S1 digestion with concomitant reduction in the supercoiled form. BND cellulose chromatographic studies also suggested the formation of single strandedness and/or distortions in the alkali treated DNA molecule. Antisera raised against the alkali treated DNA exhibited high cross-reactivity with both single stranded and Z DNA. Moreover, a significant reduction in transformation frequency of the treated DNA molecule compared with the untreated control further ascertained the structural alterations in DNA as a result of exposure to mild alkali.
已使用S1核酸酶水解、苯甲酰化萘甲酰化二乙氨基乙基纤维素（BND-纤维素）色谱法以及某些免疫学和遗传学技术来评估弱碱（pH10.0）对DNA分子的影响。与未处理的对照相比，天然小牛胸腺DNA在暴露于碱性pH10.0后，经S1核酸酶水解会释放出大量酸溶性核苷酸。对于pBR322 DNA，线性DNA种类的数量在S1消化时增加，同时超螺旋形式减少。BND纤维素色谱研究也表明，经碱处理的DNA分子形成了单链和/或扭曲。针对经碱处理的DNA产生的抗血清与单链DNA和Z-DNA均表现出高交叉反应性。此外，与未处理的对照相比，经处理的DNA分子的转化频率显著降低，这进一步确定了由于暴露于弱碱而导致的DNA结构改变。
