Rapid communication: mapping of the Mannose-Binding Lectin 2 (MBL2) gene to pig chromosome 14.

Name of Gene Marker. MBL2, Mannose-Binding Lectin 2 (Mannose-Binding Protein, Serum; MBP1). Source and Description of Primers. Primers (set A) were designed based on human (GenBank accession number X15422) and cattle (GenBank accession number D73408) MBL2 sequences available in the GenBank database. Using porcine genomic DNA as a template in the PCR, these primers amplified an 831-bp fragment. Set B primers were designed based on pig DNA sequence data generated using set A primers. This latter 771-bp pig sequence has been deposited into GenBank as a sequence tagged site (STS), accession number AF208528. Primer Sequences. Set A primers were as follows: forward 5′ AGG CAA AGA TGG GCG TGA TGG 3′ and reverse 5′ ACC AGG GTC TCC TTT TTG GCC 3′. Set B primers were as follows: forward 5′ AAG GGA GAA CCA GGT ATA GG 3′ and reverse 5′ AAT ATT TCC TGG AGG TCC C 3′. Method of Detection. The polymerase chain reaction with set A primers was performed in 10L reactions including 0.38 U Taq Gold DNA polymerase (PerkinElmer, Branchburg, NJ), 1×PCR buffer, 2.0 mM MgCl2, 0.2 mM of each dNTP, 3 pmol of each primer, and 12.5 ng genomic DNA. Thermal cycling was carried out in a Robocycler (Stratagene, La Jolla, CA) or a MJ Research, PTC-100 instrument (Watertown, MA) and included initial denaturation at 95°C for 5 min, followed by 35 cycles at 95°C for 45 s, 56°C for 1 min, 72°C for 1.5 min, and a final extension step at 72°C for 5 min. Products from Yorkshire, Landrace, and Meishan breeds were directly sequenced using dye terminators and an ABI 377 sequencer (Perkin-Elmer, Foster City, CA). The set B primers were used to amplify a 749-bp fragment. This product was used for physical and linkage mapping. Set B-PCR was performed using the MJ Research, PTC-100 instrument and conditions as for set A except that the MgCl2 concentration was 1.5 mM, Taq DNA polymerase (Promega, Madison, WI) was used, and the first denaturation step in the PCR was