Construction of a cytogenetically characterized porcine somatic cell hybrid panel and its use as a mapping tool

Abstract

A new panel of cytogenetically characterized pig-rodent somatic cell hybrids was constructed and tested for twelve microsatellite markers with PCR. Cytogenetic characterization of hybrids was accomplished by fluorescence painting and GTG-banding of metaphase chromosomes. The panel consists of 15 independent pig-hamster and 6 independent pig-mouse cell lines. In the panel, all pig autosomes and the X Chromosome (Chr) are represented, and it is informative for all chromosome pairs except 2-14, 2-15, 3-9, 14-15, 14-16, and 16-17. The microsatellites tested were S0022, S0023, S0084, S0098, S0112, S0113, S0114, S0115, S0117, S0118, S0119, and S0120. The PCR results obtained in the 21 hybrids were compared with the cytogenetic data and analyzed for concordancy and correlation. Eight microsatellites could be assigned to specific pig chromosomes, confirming seven assignments based on linkage analysis.

Mapping of the Na+, K+-ATPase subunit α 2 (ATP1A2) and muscle phosphofructokinase (PFKM) genes in pig by somatic cell hybrid analysis

“…Using a panel of 21 pig±rodent somatic cell hybrids (Zijlstra et al 1996), we have mapped the porcine Na + , K + -ATPase subunit a 2 (ATP1A2) and muscle phosphofructokinase (PFKM) genes. PCR primers were designed from the sequences Z98783 (ATP1A2 primers: forward 59-CAC GGT CAA TAA ATC TCA AAT G-39, reverse 59-ACC CTA AGG AAA ATG GAA GAC-39) and X94254 (PFKM primers: forward 59-CAC AGT GAC CAG TTG GCA TT-39, reverse 59-TAT GCT GAA GCC ATG GTC AG-39).…”

“…Concordancy percentages and correlation coefficient (w) values(Chevalet & Corpet 1986) for ATP1A2 and PFKM PCR products and the presence or absence of individual pig chromosomes in a pig±rodent somatic cell hybrid panel(Zijlstra et al 1996) …”

Contribution of Radiation Hybrids to Genome Mapping in Domestic Animals

“…In other animals, mapping studies began 10 years later, and somatic cell hybrids have also proved to be a useful tool for gene mapping. Somatic cell hybrid panels were developed for cattle [Heuertz and Hors-Cayla, 1978;Womack and Moll, 1986], dog [Bruns et al, 1978], pig [Gellin et al, 1981;Rettenberger et al, 1994;Zijlstra et al, 1996], rabbit [Echard et al, 1981], cat [O'Brien and Nash, 1982], sheep [Saïdi-Mehtar et al, 1981;Tucker et al, 1981;Burkin et al, 1998], river buffalo [Rank et al, 1998], and horse [Shiue et al, 1999]. However, in most cases the mapping power of the panel remained weak due to the large size of the chromosome fragments retained in the hybrid cells (often whole chromosomes).…”

Construction of a whole-genome radiation hybrid panel for high-resolution gene mapping in pigs

“…Three somatic cell hybrid panels have been produced in swine (Rettenberger et al, 1994;Yerle et al, 1996;Zijlstra et al,. 1996) and have been used to map porcine genes (Rettenberger et al 1996;Messer et al 1997;Sun et al, 1997;TosserKlopp et al, 1998), as well as ESTs (Fridolfsson et al 1997;Jorgensen et al, 1997;Clouscard-Martinato et al, 1998;Winterö et al, 1998) or microsatellites .…”