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.

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