Structural Analysis of Dna Complementary to Bovine Parathyroid Hormone Messenger-Rna and Restriction Endonuclease Mapping of the Parathyroid Hormone Gene

To facilitate studies on the structure and regulated expression of the gene for parathyroid hormone (PTH), full length single and double stranded DNA complementary (cDNA) to bovine PTH mRNA was synthesized by reverse transcriptase and E. coli DNA polymerase I. Single stranded cDNA migrated as a single discrete band of 750 nucleotides by denaturing gel electrophoresis. Hybridization of the cDNA to excess template mRNA revealed one rapidly hybridizing component consisting of 50% of the cDNA. Although second strand cDNA synthesis yielded incomplete transcripts at 15(DEGREES)C, incubations between 20-37(DEGREES)C produced one major cDNA species of 1500 nucleotides with a hairpin loop. The loop was specifically cleaved with S1 nuclease to produce a 700bp double stranded species. A restriction map was constructed of the population of cDNA molecules. Fragments corresponding to the 5' terminus of the sense strand were identified by comparing the fragment sizes obtained before and after cleavage of the hairpin loop.

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

• Gordon, David Frederick

• Biology, Animal Physiology

Identifier

(UMI)AAI8218475

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/71416