Title page for ETD etd-02162010-020558


Type of Document Master's Thesis
Author Ko, Thong-Sung
URN etd-02162010-020558
Title Development and characterization of cell-free amino acid incorporation systems from Cellvibrio gilvus
Degree Master of Science
Department Biochemistry and Nutrition
Advisory Committee
Advisor Name Title
Barnett, Lewis B. Committee Chair
Ackerman, C. J. Committee Member
Bunce, George Edwin Committee Member
Campbell, T. Colin Committee Member
King, Kendall W. Committee Member
Keywords
  • amino acids
Date of Defense 1967-02-15
Availability restricted
Abstract

A ribosomal system has been derived from Cellvibrio gilus which is active with respect to amino acid incorporation into hot trichloroacetic acid-insoluble material. Optimal incorporation systems showed a requirement for ribosomes, pH 5 enzyme protein, ATP plus ATP-generating system, magnesium ion, ammonium ion, and GTP. 14C-phenylalanine incorporation was stimulated by poly U in the presence of t-RNA which has been derived from this bacterium. The t-RNA had some activation effect on poly U-directed sup>14C-phenylalanine incorporation, whereas no effect on natural m-RNA primed sup>14C-leucine incorporation was observed. A complementary unlabeled amino acid mixture inhibited sup>14C-leucine incorporation. sup>14C-leucine incorporation was greatly inhibited by RBAase and puromycin, moderately inhibited by chloramphenicol, but not effected significantly by DNAase and actinomycin D. The optimum pH and temperature for sup>14C-leucine incorporation were 7.8 and 300° C, respectively. Polyamiaes (putrescine, spermine, and spermidine) had a slight stimulatory effect on poly U-directed.14C-phenylalanine incorporation, but no effect on 14C-Leucine incorporation.

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