Dissertations from ProQuest

Title

Isolation and characterization of the rat liver connexin-32 gene

1990

Article

Degree Name

Doctor of Philosophy (Ph.D.)

Department

Biochemistry and Molecular Biology

First Committee Member

R. K. Werner - Committee Chair

Abstract

The gene for connexin-32, a protein component of rat liver gap junctions, has been isolated. The clone for this gene was independently isolated several times with oligonucleotide and cDNA probes. Other genomic clones with homology to the oligonucleotide and unknown transcripts from various rat tissues were also isolated.The connexin-32 gene was extensively characterized by restriction mapping, DNA sequencing, and transcript mapping. Strong evidence that this clone represents the authentic connexin-32 gene was obtained from Southern blots of rat genomic DNA probed with the connexin-32 cDNA clone. Fragment sizes homologous to the cDNA were equal to those predicted by the map of the genomic clone.This gene has several interesting characteristics. A 6 kb intron exists in the 5$\sp\prime$ untranslated portion of the transcript. The gene lacks a typical TATA element and has at least three areas of transcription start sites. Sequence elements with homology to transcription factor SP1 binding sites were observed in the promoter. Other elements with homology to the sequence required for transcriptional induction by cAMP were found in the promoter and the intron. This gene has features common to "housekeeping" genes and its isolation permits further study of the promoter in relation to the biosynthesis of gap junctions.

Keywords

Biology, Molecular; Chemistry, Biochemistry