Biochemistry, Biophysics, and Structural Biology Commons^™

A Temperature-Sensitive Mutant Of Escherichia Coli Affected In The Alpha Subunit Of Rna Polymerase, Majid Mehrpouyan

Electronic Theses and Dissertations

A temperature-sensitive mutant of Escherichia coli affected in the alpha subunit of RNA polymerase has been investigated. Gene mapping and complementation experiments placed the mutation to temperature-sensitivity within the alpha operon at 72 min on the bacterial chromosome. The rate of RNA synthesis in vivo and the accumulation of ribosomal RNA were significantly reduced in the mutant at 44$\sp\circ$C. The thermostability at 44$\sp\circ$C of the purified holoenzyme from mutant cells was about 20% of that of the normal enzyme. Assays with T7 DNA as a template showed that the fraction of active enzyme competent for transcription was reduced as a …

The Use Of Synthetic Oligodeoxyribonucleotides In Direct Analysis Of Single Nucleotide Variations And Inhibition Of Eukaryotic Gene Expression, Daniel Y. Wu

Loma Linda University Electronic Theses, Dissertations & Projects

The present thesis work contributes to two areas of molecular genetics, namely the development of methods for identifying single nucleotides and the understanding of the mechanism by which antisense oligonucleotides inhibit eukaryotic gene expression. Three single nucleotide detection methods are presented. The first method utilizes hybridization with synthetic oligonucleotide probes to detect single or multiple nucleotide differences between alleles in genomic DNA without electrophoretic separation. Total genomic DNA (either digested or undigested with restriction endonucleases) was immobilized in depressions in an agarose gel (in situ dots) and hybridized with radiolabeled, allele-specific oligonucleotide probes. We show that this method can be …

Nucleotide Sequence And Further Characterization Of The Synechococcus Sp. Strain Pcc 7002 Reca Gene: Complementation Of A Cyanobacterial Reca Mutation By The Escherichia Coli Reca Gene, R C. Murphy, G E. Gasparich, D A. Bryant, R D. Porter

Gail Gasparich