Isolation of mutants lacking branched-chain amino acid transaminase

Variants of the Chinese hamster ovary cell have been isolated which can no longer grow when valine, leucine, or isoleucine is replaced in the culture medium by its respective -keto acid: -ketoisovaleric acid, -ketoisocaproic acid, or -keto--methylvaleric acid. These variants lack branched-chain amino acid transaminase activity. Evidence is presented indicating these variants to be single gene mutants. Genetic evidence is also presented confirming previous biochemical evidence that a single enzyme carries out transaminase functions on valine, leucine, and isoleucine. The branched-chain transaminase-deficient (trans^–)mutants can be reverted to wild-type behavior by treatment with mutagenic agents. These mutants promise to be useful in exploring regulatory mechanisms in biochemical, genetic, and cancer research.Trainee, U.S. Public Health Service Grant No. GM00781-17.