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

Abstract

We have developed a panel of 152 whole-genome radiation hybrids by fusing irradiated diploid pig lymphocytes or fibroblasts with recipient hamster permanent cells. The number and size of the porcine chromosome fragments retained in each hybrid clone were checked by fluorescence in situ hybridization with a SINE probe or by primed in situ labeling (PRINS) with SINE-specific primers. A strategy based on the interspersed repetitive sequence polymerase chain reaction (IRS-PCR) was developed for selected clones to determine if the large fragments painted by the SINE probe corresponded to one pig chromosome or to different fragments of several chromosomes. This strategy was buttressed by a double PRINS approach using primers specific for α-satellite sequences of two different groups of swine chromosomes. Genome retention frequency was estimated for each clone by PCR with 32 markers localized on different porcine chromosomes. Of the 152 hybrids produced, 126 were selected on the basis of cytogenetic content and chromosome retention frequency to construct a radiation hybrid map of swine chromosome 8. Our initial results for this chromosome indicate that the resolution of the radiation hybrid map is 18 times higher than that obtained by linkage analysis.

ChickRH6: a chicken whole-genome radiation hybrid panel

“…The method was adapted from the one first described by Walter et al [36] and widely used [22,38]. The chicken donor cells used for constructing this panel were normal diploid fibroblasts.…”

“…WGRH panels are now available in many species including human [13,27,33], mouse [23], rat [22], dog [35], cat [26], cow [37], pig [38] and horse [17]. Zebrafish WGRH panels and RH maps have been published [11,14], demonstrating that RH technology can also be used for non-mammalian vertebrates.…”

High-Resolution Radiation Hybrid Map of Wheat Chromosome 1D

“…An advantage of RH mapping is that different doses of radiation can be used to construct maps with varying levels of resolution Lunetta et al 1996;Schuler et al 1996;Stewart et al 1997). This work has been followed by RH mapping in a number of other animal species (McCarthy et al 1997;Priat et al 1998;Yerle et al 1998;Hawken et al 1999;Hukriede et al 1999;Kwok et al 1999;Murphy et al 1999;Vignaux et al 1999;Watanabe et al 1999). In addition to whole-genome RH mapping, other researchers have focused on mapping specific chromosomes and chromosomal regions in different species as well (Hoover and Floros 1998;Barendse et al 2000;Ozawa et al 2000;Liu et al 2002;Rink et al 2002).…”

Polymorphisms in an obesity-related gene (PCSK1) are associated with fat deposition and production traits in Italian heavy pigs

“…The INRA-Minnesota 7000 rads RH panel (IMpRH panel; Yerle et al, 1998) consisting of 118 rodent-porcine hybrid cell lines was screened by means of PCR using primer pair indicated in Table S1. No PCR fragment was obtained from the control rodent genomic DNA.…”