Isolation, Mapping, Expression Profile Of PITPNA & PRPSAP1 Genes And Association Analysis Of PSME3 & FDXR Genes On SSC12

In this study 2 novel genes' fragment were isolated from SSC12 by expoiting information resources in pig ESTs database and comparative map, and the related study include gene mapping, expression profile and CDS analysing. The initial association between gene polymorphism and some production traits was also investgated about the other 2 genes on SSC12. The main results are as follows:1. Pig ESTs were scanned according to cDNA sequences of human homologous genes on chromosome 17. Highly conserved pig ESTs were assembled to contigs by seqMan program of DNAStar, and matched primers of the contigs were designed to isolate porcine PITPNA and PRPSAP1 gene fragments from pig genomic DNA.2. The full length CDS of PITPNA and PRPSAP1 genes were obtained through in silico cloning. 2 deduced swine amino acid sequences of these two genes show 99% and 98% high similarity to human sequences coded by the homologous genes, respectively.3. The pig x rodent somatic cell hybrid panel (SCHP) was used for regional mapping of pig PITPNA and PRPSAP1 genes. The mapping results show that PITPNA gene was localized on SSC12 q11-q15 (P=0.9711, r=0.9035) and PRPSAP1 gene is physically mapped on SSC12 p11-(2/3 p13)( P=0.8901, r=1.000), respectively.4. The INRA-University of Minnesota porcine radiation hybrid (IMpRH) panel was exploited to assign the two novel genes on precise chromosome region. The result revealed pig PITPNA gene is closely linked to marker S0106(LOD=5.99), the PRPSAP1 gene is closely linked to S0143 and SW2494, with LOD score threshold of 10,09 and 8,38, respectively.5. Expression profiles of PITPNA and PRPSAP1 genes in seven different tissues from Landrace pigs and six different tissues from Hubei White pigs were analysed by semi-quantitative RT-PCR. The result show pig PRPSAP1 gene is a little low expressed in heart, but widely expressed in other tissues and show no significant difference among tissues, pig PITPNA gene is not expressed in the adipose and show widely expressed among other tissues with insignificant difference.6. Studies on polymorphism of FDXR and PSME3 genes by PCR-RFLP revealed two polymorphic sites. The 2 polymorphic sites were genotyped in the experimental pig population (traits and DNA sample were well-documented) constructed under the cooperation of our lab and husbandry bureau of Tongcheng county. Association between the investigated genotypes and phenotypes of production traits in pigs including part of meat quality, growth and carcass traits were analyzed. The study revealed that①There are highly significant associations (P<0.01) between the genotypes of the 182^nd polymorphic site of PSME3 and the phenotype of Backfat at the rump, significant association (P＜0.05) between the genotypes and the Loin colour or Marbling score were showed too.②The difference is highly significant (P＜0.01) between GG genotype and AG genotype of the 182na polymorphic site of PSME3 in the backfat at the romp, and the difference is also highly significant (P＜0.01) between AA genotype and AG genotype in the phenotype of Marbling score. The difference between the genotypes of AA and AG in the phenotype of Backfat at the loin, AA and AG in the backfat at the rump, GG and AG in Average backfat thickness, GG and AG in Loin colour, AA and AG in Marbling score are all significant (P＜0.05) according to the study.③T here are significant associations of the genotypes of the 562nd polymorphic site of FDXR with the Loin-muscle area (LMA) (P＜0.05).