Date of Award:

5-1-1984

Document Type:

Thesis

Degree Name:

Master of Science (MS)

Department:

Biology

Department name when degree awarded

Life Sciences:Biology

Committee Chair(s)

Jon Y. Takemoto

Committee

Jon Y. Takemoto

Committee

Fred Post

Committee

John Simmons

Abstract

The purpose of this work was to construct a DNA vector containing the Escherichia coli lactose operon and devise a method for the subsequent introduction into Rhodopseudomonas sphaeroides. Expertise in methods for the purification and manipulation of DNA was developed and improvements upon methods were devised. PRSF 1010 proved to be a poor cloning vector due to a lack of selectable resistance markers and useable restriction sites. The lac operon was cloned into pBR 325 using either HindIII or BamHI restriction endonucleases. Restriction maps of the resulting plasmids were constructed. Attempts to transform R. sphaeroides with plasmid DNA by established protocols for other bacteria were unsuccessful. A procedure is outlined by which a lac operon containing plasmid can be introduced into R. sphaeroides via conjugation with E. coli.

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