Single-Stranded-Dna Sp6 and T7 Bacteriophage Promoter Plasmids for Engineering Mutant Rnas and Proteins: Extensions and Deletions at the Amino Terminus Which Alter the Activity of Preproparathyroid Hormone Signal Peptide

Protein engineering systems have been developed which simplify the cloning, oligonucleotide directed mutagenesis, enzymatic sequence analysis, and expression of mutant proteins, all from a single vector. Chimeric phage-plasmid expression vectors, or phagemids, containing the intergenic region of the single stranded DNA bacteriophage f1 and either a bacteriophage SP6 or T7 promoter have been constructed. Coinfection of E. coli with these chimeric plasmids and a single stranded DNA helper phage results in the replication and secretion of the pSP6(.)f1 or pTZ plasmids as single stranded DNA viral particles. A single species of RNA can be transcribed from these vectors due to the stringent specificity of bacteriophage T7 or SP6 RNA polymerase for their respective promoter DNA sequences. Thus, these vectors are suitable for the efficient biosynthesis of large amounts of single or double stranded DNA, and translationally active RNA.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Physiology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Mead, Alan David

• Biology, Molecular

Identifier

(UMI)AAI8701564

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/71445