Rapid communication: Linkage and physical mapping of the porcine basic fibroblast growth factor (FGF2) gene1

Genus and species. Sus scrofa. Locus. Pig Fibroblast Growth Factor 2 (FGF2) gene. Source and Description of Primers. Primers were designed from human sequence (GenBank accession no. J04513.1) to amplify a 167-bp fragment within exon 1 of FGF2 from pig genomic DNA. This fragment was identified as FGF2 with 93% homology to the human FGF2 sequence. Primer Sequences. Primers designed from human sequence were as follows: forward primer: 5′ GCA GCC GGG AGC ATC ACC AC 3′; reverse primer: 5′ TCG CTC TTC TCC CGG ACC C 3′. Pig-specific primers were as follows: forward primer: 5′ TGA ATA TAA AAA ATC CTA AGC GGT TGC ACT 3′; reverse primer 5′ CGC TCT TCT CCC GGA CCC 3′. Method of Detection. The PCR reaction using the pigspecific primers was performed using 12.5 ng of porcine genomic DNA, 1× PCR buffer, 1.5 mM MgCl2, 100 M dNTP, 2.5 pM of each primer, and 0.35 units of Taq DNA polymerase (Promega, Madison, WI) in a final volume of 10 L. The PCR program consisted of an initial 4 min 94°C denaturing, 35 cycles of 45 s at 94°C, 45 s at 57°C, 45 s at 72°C, and a final 5-min extension at 72°C in a MJ PTC-100 thermocycler (MJ Research, Watertown, MA). The PCR product was digested with BtsI at 37°C overnight and fragments were separated by electrophoresis on a 4% Nusieve gel (BMA, Rockland, ME). Description of Polymorphism. A single nucleotide polymorphism (SNP), C/T base pair change (silent mutation), was detected in the pig sequence at the human FGF2 nucleotide position 568 (GenBank accession no. J04513.1). A BtsI RFLP test was developed for this exon 1 SNP by designing pig-specific primers to amplify a 99-bp fragment and incorporate a BtsI restriction enzyme site. The BtsI digestion of the 99-bp exon 1 PCR fragment produced the following genotypes: homozy-