Radiation hybrid mapping of 273 previously unreported porcine microsatellites

The single-stranded DNA-binding protein 1 (SSBP1) is a transcription factor involved in the regulation of the thyroid stimulating hormone receptor (TSHR).It belongs to a family of SSBPs that interact with RNA and with the promoter of retroviruses, and have an important role in RNA processing.The SSBP1 maps to human chromosome 7 (HSA7).Polymerase chain reaction (PCR) primers flanking the third and fourth introns of the ovine SSBP1 gene were designed using the targeted intronic polymorphic sequence identification (TIPS) procedure 1 .The PCR products were tested for similarity with non-redundant (nr) sequences in GenBank (http://www.ncbi.nlm.nih.gov/) using the BLASTN algorithm 2 .The coding sequences flanking the introns were unequivocally aligned to their homologous sequences in the human SSBP1 gene (GenBank accession number NM_003143).The ovine SSBP1 sequence was deposited with the GenBank nucleotide sequence database under accession number AY115486.A repeated sequence (T) 6, identified within the third intron at position 469 in AY115486, showed variation in different individuals.This microsatellite was designated as OARCDT7.We designed PCR primers flanking OARCDT7 and used it to map SSBP1 in sheep. Primer sequences: OARCDT7 -Forward primer: 5¢-GAGCTTA ATGCTTGTTTTGTGG-3¢ OARCDT7 -Reverse primer: 5¢-CAGGAAACAAGTGTGAGG-AGC-3¢Analysis ⁄ Mendelian inheritance: The OARCDT7 was amplified by PCR and analysed by denaturing polyacrylamide gel electrophoresis.Two alleles were identified and their codominant segregation was verified in the AgResearch International Mapping Flock (IMF), comprising nine three-generation families with a total of 127 animals 3 . Chromosomal location:The OARCDT7 was mapped against the markers on the latest version of the sheep framework map 4 .Multipoint linkage analysis of the IMF pedigrees using CRI-MAP 5 localized OARCDT7 in the distal part of ovine chromosome 4, between markers BM2023 and CSRD219, with its most likely position at BM2023.Two-point likelihood of odds (LOD) scores and recombination fractions for these loci are shown in Table 1.References