Journal Article

ACCELERATED PAPER: Cloning, genetic mapping and expression studies of the rat <i>Brca</i>1 gene

Kai-Shun Chen, Laurie A. Shepel, Jill D. Haag, Gerlyn M. Heil and Michael N. Gould

in Carcinogenesis

Volume 17, issue 8, pages 1561-1566
Published in print August 1996 | ISSN: 0143-3334
e-ISSN: 1460-2180 | DOI:
ACCELERATED PAPER: Cloning, genetic mapping and expression studies of the rat Brca1 gene

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The breast cancer gene BRCA1 has previously been cloned from both human and mouse. We cloned a fragment of the rat Brca1 homologue in order to map it and explore its biological function. Partial cDNA fragments of the rat Brca1 homologue were isolated by RT-PCR. Sequence analysis revealed that the RING-finger domain is well conserved among rat, mouse and human. Rat Brcal mRNA was expressed in most tissues studied with the highest level in testis, consistent with studies in human and mouse. Next, intron 6-containing DNA fragments were amplified by PCR from WKY and WF strains. The splicing sites between exon 6 and exon 7 are conserved between rat and human. Partial sequencing of the rat Brca1 intron 6 revealed a polymorphism of a pentanucleotide TTTTG repeat between the WKY and WF strains. With this intragenic microsatellite marker, we were able to map precisely the rat Brcal gene to chromosome 10 using a genetic linkage study of (WKY×WF)F1×WF baccross rats. Brca 1 cosegregates with marker BAND3A, and is flanked by R5123 and R5842. Using this polymorphic marker, we also investigated the loss of heterozygosity (LOH) of the Brcal microsatellite marker in carcinogen- or radiation-induced mammary carcinomas in (WF×F344)F1 female rats. No LOH or somatic microsatellite instability was detected in 18 DMBA-induced tumors studied. Only one LOH of the F344 allele was observed in 26 radiation-induced tumors tested. Ribon-uclease protection assays demonstrated that Brca1 mRNA levels are similar in normal rat mammary glands and mammary carcinomas of various etiologies, including those induced by DMBA, NMU, activated-neu and activated-ras oncogenes.

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Subjects: Clinical Cytogenetics and Molecular Genetics

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