Narayanan K R
Arch Toxicol. 1977 Sep 21;38(1-2):61-73. doi: 10.1007/BF00293664.
Several bioanalytical techniques including electrofocusing, two-dimensional technique comprising electrofocusing and electrophoresis, and chromatography have been evaluated and improved to provide a method for detecting gene mutations in mice. The use of 1 mm gels for both electrofocusing and electrophoresis in the two-dimensional technique resulted in better resolution, shorter separation time, and reduced costs. An electrophoretic apparatus for the second direction electrophoresis has been designed which eliminated cutting of a flat gel into strips prior to electrophoresis. These changes reduced the total time for a two-dimensional separation to 4 h. The scope and application of isoelectric focusing in detecting gene mutations in mice have been improved by the agar contact replica method. This method allows the simultaneous analysis of several genetic loci besides facilitating automation of evaluation, documentation, and storage of zymograms. Preliminary studies on the detection of heritable metabolic disorders of amino acid metabolism in mice show a potential for such studies in estimating mutation rates in mice. Some possible techniques for such studies and their implications are discussed.
包括电聚焦、由电聚焦和电泳组成的二维技术以及色谱法在内的几种生物分析技术已得到评估和改进，以提供一种检测小鼠基因突变的方法。在二维技术中，电聚焦和电泳均使用1毫米凝胶，从而实现了更高的分辨率、更短的分离时间并降低了成本。已设计出一种用于第二方向电泳的电泳装置，该装置无需在电泳前将扁平凝胶切成条带。这些改进将二维分离的总时间缩短至4小时。琼脂接触复制法提高了等电聚焦在检测小鼠基因突变方面的范围和应用。该方法除了便于酶谱的评估、记录和存储自动化外，还能同时分析多个基因位点。对小鼠遗传性氨基酸代谢紊乱检测的初步研究表明，此类研究在估计小鼠突变率方面具有潜力。文中讨论了一些用于此类研究的可能技术及其意义。
