Development and assignment of bovine-specific PCR systems for the Texas nomenclature marker genes and isolation of homologous BAC probes

Abstract

-In 1996, Popescu et al. published the Texas standard nomenclature of the bovine karyotype in which 31 marker genes, already mapped in man, were chosen to permit unambiguous identification and numbering of each bovine chromosome. However, specific PCR systems were not available for each marker gene thus preventing the assignment of part of these markers by somatic cell hybrid analysis. In addition, some difficulties remained with the nomenclature of BTA25, BTA27 and BTA29. In this work, specific PCR systems were developed for each of the marker genes except VIL1 (see results), from either existing bovine or human sequences, and a bovine BAC library was screened to obtain the corresponding BAC clones. These PCR systems were used successfully to confirm the assignment of each marker gene (except for LDHA, see results) by analysis on the INRA hamster-bovine somatic cell hybrid panel. The difficulties observed for LDHA and VIL1 are probably due to the fact that these genes belong to large gene families and therefore suggest that they may not be the most appropriate markers for a standardisation effort. This panel of BACs is available to the scientific community and has served as a basis for the establishment of a revised standard nomenclature of bovine chromosomes. bovine / BAC library / cytogenetics / mapping / Texas standard

Construction and characterization of a bovine BAC library with four genome-equivalent coverage

“…Only one microsatellite was not found in the library. The library was also successfully screened for the 31 marker genes of the Texas standard nomenclature of the bovine karyotype [4], providing an accurate set of BAC clones which were used to resolve chromosome identification problems [4,6]. Fluorescence in situ hybridisation was performed as described [6] for over 50 BAC clones: none of the clones was chimeric.…”

Assignment¹ of the bovine <i>PTGFR</i> and <i>HSD3B1</i> genes to bovine chromosome 3 with somatic and radiation hybrid panel mapping

“…The PCR reactions were performed in 10-l reaction mixtures 1 To our knowledge this is the fi rst time PTGFR has been mapped in cattle. This is a more precise localization of HSD3B1 previously mapped to BTA3q12 by Gautier et al, 2001.…”

“…PTGFR was mapped for the fi rst time. HSD3B1 was previously assigned to BTA3 by Gautier et al (2001), by analysis on the INRA hamster-bovine somatic cell hybrid panel. In this report, physical mapping of HSD3B1 was also performed by PCR typing on a different bovine-hamster somatic cell panel with a different PCR primer pair, followed by RH mapping.…”

A Case of Trisomy 28 in Cattle Revealed by both Banding and FISH-Mapping Techniques

“…Drops of cell suspension obtained from BrdU-treated blood cell cultures (CHRISTENSEN and JUUL 1999) kept frozen until now, were spread on slides which were used for both RBG-banding (R-bands by late incorporation of 5-Bromodeoxyuridine and Giemsa staining) and fluorescence in situ hybridization (FISH) techniques. As probes, the bovine BAC-clones mapping LTF (lactotransferrin, marker of BTA22) and CGNl (conglutinin 1, marker of BTA28) (TEXAS NOMENCLA-TURE 1996;ISCNDB2000 2001GAUTIER et al 2001) were used. Slide-treatment for FISH-technique, in situ hybridization, signal detection, image capturing by CCD-camera, image processing and printing were as recently reported (IANNUZZI et al 2000).…”

“…Since specific type I molecular markers have been assigned to each cattle chromosome (TEXAS NOMEN-CLATURE 1996;GAUTIER et al 2001) and new Q-, Gand R-banded karyotypes have recently been arranged (ISCNDB2000 200 1) using the Texas system (1 996) as point of reference, the use of specific markers may be very useful in clinical cytogenetics.…”