Characterization of reciprocal translocations in pigs using dual-colour chromosome painting and primed in situ DNA labelling.
We report the use of dual-colour chromosome painting to determine the exact nature of certain chromosome rearrangements observed in the pig (Sus scrofa domestica). The chromosomal abnormalities were detected by GTG- and RBG-banding techniques. The initially proposed interpretations were: (1) rcp(6;13)(p1.5;q4.1); (2) rcp(11;16)(p1.4;q1.4); (3) rcp(6;16)(p1.1;q1.1); (4) rcp(13;17)(q4.1;q1.1); (5) rcp(6;14)(q2.7;q2.1); (6) rcp(3;5)(p1.3;q2.3); (7) rcp(2; 14)(q1.3;q2.7); (8) rcp(15;17)(q1.3;q2.1). Hybridizations were carried out with biotin- and digoxigenin-labelled probes obtained by priming authorizing random mismatches polymerase chain reaction (PARM-PCR) amplification of porcine flow-sorted chromosomes. In some cases, i.e. (1), (4), (5), (6), (7) and (8), the fluorescence in situ hybridization (FISH) results allowed confirmation of the interpretations proposed with classical cytogenetic methods. Chromosome painting proved the reciprocity of the translocation in cases (1), (6) and (8), whereas modifications of the formula were proposed for case (2). Primed in situ DNA labelling (PRINS) experiments have also been carried out in case (3) using a primer specific for the centromeres of acrocentric chromosomes (first experiment) or a primer specific for the centromeres of a subset of meta- and submetacentric chromosomes including chromosome 6 (second experiment). It allowed us to demonstrate that the breakpoints occurred in the centromeric region of chromosome 16 and in the p. arm of chromosome 6, just above the centromere.
Pinton
A
A
ENVT, Laboratoire de Cytogénétique, Toulouse, France.
Ducos
A
A
Séguéla
A
A
Berland
H M
HM
Darré
R
R
Darré
A
A
Pinton
P
P
Schmitz
A
A
Cribiu
E P
EP
Yerle
M
M
D000818:Animals; D001710:Biotin; D020223:Chromosome Painting; D002875:Chromosomes / Q000737:chemistry; D004247:DNA / Q000032:analysis; D017931:DNA Primers; D004076:Digoxigenin; D017404:In Situ Hybridization, Fluorescence; D016133:Polymerase Chain Reaction / Q000379:methods; D013552:Swine / Q000235:genetics*; D014178:Translocation, Genetic
D017931:DNA Primers; D001710:Biotin; D004247:DNA; D004076:Digoxigenin