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Genetic heterogeneity and selection signature at the KIT gene in pigs showing different coat colours and patterns£¨ÌâÄ¿£© Sequencing of the KIT gene The complete coding region of the KIT gene was sequenced in 35 pigs of different breeds: 10 Nero Siciliano pigs, four with completely black coat colour, four with white spots but without duplication of the KIT gene, two with white spots but with the duplication of the KIT gene; three grey animals (one was homozygous for intron 18 deletion and two were heterozygous at this polymorphic site); six Cinta Senese; four Hampshire; four Duroc, two Italian Large White, one carrying the splice mutation and one not carrying this mutation; two Pietrain, one positive and one negative at the duplication test; two Meishan; two wild boars. PCR primers were designed with PRIMER3 (https://fokker.wi.mit.edu/ primer3/input.htm) using pig genomic sequence AC141857 (clone RP44-473N18) containing the 21 KIT exons. BLASTN (bl2seq) analysis (https://blast.ncbi.nlm.nih.gov/Blast.cgi) with cDNA sequences of the porcine KIT gene was used to identify exon borders in the genomic sequence. PCR primers were placed in the intronic region except, for the last exon, which contains a long 3 untranslated region (UTR). PCR primers and amplification conditions are reported in Table S1. Sequences were obtained as described in Table S1. Sequences were assembled and analysed and polymorphisms were detected using CodonCode Aligner (https:// www.codoncode.com/aligner), with the AC141857 sequence included as a reference. All sequence chromatograms were visually inspected. ÊÖ»ú±à¼£¬Ï£ÍûÓиßÊÖ°ï·Òëһϣ¬ÏÈллÁË£¡ [ Last edited by yzwan1 on 2011-10-28 at 19:07 ] |
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