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Detection of human somatic cell structural gene mutations by two‐dimensional electrophoresis
Author(s) -
Hanash S.M.,
Chu E.H.Y.,
Kuick R.,
Skolnick M.,
Neel J.,
Strahler J.,
Pivirotto S.,
Niezgoda W.
Publication year - 1987
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.340020103
Subject(s) - mutant , somatic cell , microbiology and biotechnology , mutagenesis , gene , biology , ethylnitrosourea , gel electrophoresis , mutation , spots , structural gene , cell culture , genetics , botany
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.