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小鼠染色体特异性DNA文库的构建及其在检测X射线诱导畸变中的应用。

Construction of mouse chromosome-specific DNA libraries and their use for the detection of X-ray-induced aberrations.

作者信息

Boei J J, Balajee A S, de Boer P, Rens W, Aten J A, Mullenders L H, Natarajan A T

机构信息

Department of Radiation Genetics and Chemical Mutagenesis, State University of Leiden, The Netherlands.

出版信息

Int J Radiat Biol. 1994 May;65(5):583-90. doi: 10.1080/09553009414550671.

Abstract

We describe here the development of mouse chromosome-specific DNA libraries and their use in the detection of radiation-induced chromosome aberrations by fluorescence in situ hybridization. Large metacentric chromosomes, resulting from a translocation involving chromosomes 1, 11 and 13, were flow-sorted. Using a slit-scan technique for morphometric analysis, metacentric chromosomes were separated from normal acrocentric chromosomes and their aggregates. DNA from the metacentric chromosomes was amplified by PCR using the linker/adaptor method. In this pilot study, mouse was whole-body irradiated with 1, 2 and 3 Gy and aberrations were scored in metaphase spreads of splenocytes cultured in vitro. The results indicate that directly after radiation exposure, stable and unstable aberrations are induced at about equal frequencies in the splenocytes. The availability of chromosome-specific probes for mouse may prove very useful when analysing the behaviour of stable aberrations, as well as the testing of many suspected mutagenic carcinogens and aneugens in vivo for induction of chromosomal translocations and non-disjunction, respectively.

摘要

我们在此描述小鼠染色体特异性DNA文库的构建及其在通过荧光原位杂交检测辐射诱导的染色体畸变中的应用。对由涉及1号、11号和13号染色体的易位产生的大型中着丝粒染色体进行了流式分选。使用狭缝扫描技术进行形态计量分析,从中将中着丝粒染色体与正常近端着丝粒染色体及其聚集体分离。使用接头/衔接子方法通过PCR扩增中着丝粒染色体的DNA。在这项初步研究中,对小鼠进行全身1、2和3 Gy照射,并对体外培养的脾细胞中期分裂相中出现的畸变进行评分。结果表明,辐射暴露后直接观察到,脾细胞中稳定和不稳定畸变的诱导频率大致相等。当分析稳定畸变的行为,以及分别在体内测试许多疑似诱变致癌物和非整倍体剂诱导染色体易位和不分离的情况时,小鼠染色体特异性探针的可用性可能非常有用。

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