Construction of a porcine YAC library and mapping of the cardiac muscle ryanodine receptor gene to Chromosome 14q22?q23

Abstract

Large-scale physical mapping of the porcine genome has been limited because up to now no suitable genomic libraries for this purpose have been available. Therefore, we have constructed a yeast artificial chromosome (YAC) library from porcine lymphocytes. The library was cloned in the amplifiable vector pCGS966. A total of 10080 YAC clones was obtained and has been ordered into 105 96-well microtiter plates. An average insert size of 300 kb was calculated from the analysis of 78 randomly selected clones, giving a one-fold coverage of the porcine genome. To analyze the complexity, we have screened the library for five different genes and isolated four different clones containing parts of three of these genes. One YAC clone harboring parts of the porcine cardiac muscle ryanodine receptor (RYR2) gene allowed us to assign this locus to Chromosome (Chr) 14q22-q23. The data were confirmed by PCR analysis of a rodent-porcine hybrid cell panel.

Livestock genomics comes of age.

“…Schoeberlein, unpubl. ), sheep (Broom and Hill 1994), pig (Leeb et al 1995; S. Meier-Ewert, unpubl. ; C. Beattie, pers.…”

Modern genome research and DNA diagnostics in domestic animals in the light of classical breeding techniques

“…for further screening [73]. (iv) Large insert libraries are being established in cattle, sheep 1741 and the pig [75]. Furthermore, novel methods will be developed as genome mismatch scanning and representional difference analysis, allowing rapid scanning of whole genomes for differences [76].…”

Isolation and assignment<footref rid="foot01">¹</footref> of the UDP-glucose pyrophosphorylase gene (UGP2) to porcine chromosome 3q21→q22 by FISH and by analysis of somatic cell and radiation hybrid panels

“…On screening a porcine YAC library (Leeb et al, 1995) with the porcine UGP2 primers described above, a positive clone, LMUBy911F0441, was isolated. The 280-kb insert of this clone was directly labeled with rhodamine by using FluorRed (Amersham) for DOP-PCR (Telenius et al, 1992) and then hybridized at 42°C overnight to porcine male metaphase spreads at F10 ng/Ìl hybridization mix consisting of 50 % formamide, 2 × SSC, 10 % dextran sulfate, 600 ng/Ìl sheared herring sperm DNA (Boehringer) and 600 ng/Ìl pig Cot1 DNA.…”