Date of Award:
5-1-1987
Document Type:
Thesis
Degree Name:
Master of Science (MS)
Department:
Biology
Department name when degree awarded
Biology
Committee Chair(s)
Jon Y. Takemoto
Committee
Jon Y. Takemoto
Committee
Frederick J. Post
Committee
Raymond T. Sanders
Abstract
The purpose of this work was to attempt to synthesize, in vitro, the B875 light harvesting proteins of Rhodobacter sphaeroides from a PBR322 derived recombinant plasmid named pSS2. Large scale plasmid purification of both the recombinant plasmid and pBR322 was performed. Restriction digest patterns indicated that the extracted plasmids were the correct size and construction. An in vitro DNA directed-coupled transcription-translation protein synthesis was developed from extracts of either R. sphaeroides or Escherichia coli. Trichloroacetic acid (TCA) protein precipitation techniques were used to determine the protein synthesis activities of the extracts and plasmids. The concentration optimum for magnesium was determined for the synthesis mixtures. The in vitro synthesized polypeptides were isolated by immune-precipitation and visualized using polyacrylamide gel electrophoresis and autoradiography. The pSS2 plasmid did not synthesize the B875 polypeptides in the above synthesis systems.
Recommended Citation
Seeley, Kevin, "Attempts at In Vitro Synthesis of the B875 Light Harvesting Polypeptides of Rhodobacter sphaeroides Using Recombinant Escherichia coli Plasmid pSS2" (1987). Biology. 553.
https://digitalcommons.usu.edu/etd_biology/553
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