通过二维电泳检测人类体细胞结构基因突变
Detection of human somatic cell structural gene mutations by two-dimensional electrophoresis.
作者信息
Hanash S M, Chu E H, Kuick R, Skolnick M, Neel J, Strahler J, Pivirotto S, Niezgoda W
机构信息
Department of Pediatrics, University of Michigan, Ann Arbor 48109.
出版信息
Proteins. 1987;2(1):13-9. doi: 10.1002/prot.340020103.
The feasibility of detecting human somatic structural gene mutations by two dimensional electrophoresis has been investigated. A lymphoblastoid cell line was grown as a mass culture in the presence of ethylnitrosourea, after which cells were regrown as single cell clones. A total of 257 polypeptide spots were analyzed in gels derived from 186 clones. Four structural mutations were detected by visual analysis of the gels. Computer analysis of gels corresponding to the mutant clones was also undertaken. At a spot size threshold of 200 spots to be matched using a computer algorithm, all four mutant polypeptides were detected. These results indicate the usefulness of the two-dimensional approach for mutagenesis studies at the protein level.
已经研究了通过二维电泳检测人类体细胞结构基因突变的可行性。在存在乙基亚硝基脲的情况下，将淋巴母细胞样细胞系作为大量培养物进行培养，之后将细胞重新培养为单细胞克隆。对来自186个克隆的凝胶中的总共257个多肽斑点进行了分析。通过对凝胶的视觉分析检测到四个结构突变。还对与突变克隆相对应的凝胶进行了计算机分析。在使用计算机算法匹配的斑点大小阈值为200个斑点时，检测到了所有四个突变多肽。这些结果表明二维方法在蛋白质水平诱变研究中的有用性。
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