RGD Reference Report - Two transcripts encode rat cytochrome b5 reductase. - Rat Genome Database

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Two transcripts encode rat cytochrome b5 reductase.

Authors: Pietrini, G  Carrera, P  Borgese, N 
Citation: Pietrini G, etal., Proc Natl Acad Sci U S A 1988 Oct;85(19):7246-50.
RGD ID: 728559
Pubmed: PMID:3174630   (View Abstract at PubMed)
PMCID: PMC282162   (View Article at PubMed Central)

A cDNA expression library in lambda gt11 was screened with affinity-purified polyclonal anti-rat cytochrome b5 reductase antibodies. One positive clone out of 450,000 clones was isolated and found to be incomplete. This clone was used to rescreen the library, and a second, overlapping clone that contained the entire coding sequence was isolated. RNA gel blots showed that the two overlapping clones contained approximately 90% of the reductase mRNA sequence. Sequencing data showed (i) that rat reductase has a 93% sequence similarity with bovine and human reductase and (ii) that reductase is not synthesized as a high molecular weight precursor. Results of Southern blot analysis were consistent with the hypothesis that a single gene codes for the soluble and membrane-bound (microsomal and mitochondrial) forms of the reductase, present in erythrocytes and liver, respectively. The cloned cDNA was used to study reductase transcripts in liver and reticulocytes. Two antisense RNA probes that together covered the entire coding region and part of the noncoding region of reductase mRNA were used in RNase A protection experiments. These probes detected only one transcript in liver, suggesting that endoplasmic reticulum and mitochondrial reductase are translated from the same mRNA. In contrast, two transcripts were detected in reticulocytes, one of which mismatched the liver probe approximately 30 nucleotides downstream from the initiation codon. Since the soluble and membrane form of the reductase are known to differ at the N terminus, we suggest that this second transcript encodes soluble reductase.

Objects referenced in this article
Gene Cyb5r3 cytochrome b5 reductase 3 Rattus norvegicus

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