Assignment of porcine cyclin-dependent kinase 4 (CDK4) and oncogene c-mos (MOS) by nonradioactive nonfluorescence in situ hybridization.
Two pig genes, cyclin-dependent kinase 4 (CDK4) and the oncogene c-mos (MOS) were mapped by means of nonradioactive nonfluorescence in situ hybridization. Our approach was based on the detection of hybridized biotinylated probe by peroxidase conjugated extravidin and the reaction of peroxidase with its substrate diaminobenzidine (DAB) resulting in a dark precipitate. To increase the sensitivity of the method in single-copy gene mapping, two amplifications of the peroxidase signal were used: immunological amplification by biotinylated antiavidin, and peroxidase-catalysed deposition of biotinylated tyramide. Using this method, two 2-kb-long probes for the porcine genes CDK4 and MOS were mapped to pig chromosomes 5p12 and 4q14-15, respectively. Non-radioactive nonfluorescence in situ hybridization described here is a method of choice for gene mapping of short probes.
Musilová
P
P
Veterinary Research Institute, Hudcova, Brno, Czech Republic.
Kubícková
S
S
Rubes
J
J
D015100:3,3'-Diaminobenzidine / Q000378:metabolism; D000818:Animals; D002874:Chromosome Mapping; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases / Q000235:genetics*; D016258:Genes, mos; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009195:Peroxidase / Q000378:metabolism; D011518:Proto-Oncogene Proteins; D013552:Swine / Q000235:genetics*
D011518:Proto-Oncogene Proteins; D015100:3,3'-Diaminobenzidine; D009195:Peroxidase; D051358:Cyclin-Dependent Kinase 4; D018844:Cyclin-Dependent Kinases