| Nowadays, the production performance of pig has been improved by modern breeding technie. However, the more standard of living enhance, the more demand for quality of pork: good taste and few drug retention. So, pig breeding has two foci: one is improving lean growth rate and meat quality, the other is increasing the disease resistance to reducing growth cost. Only by excellent quality, the pig breeding industry of China can compete with other countries for world market.In this research, the porcine 20S proteasome related new genes were isolated and mapped by combining bioinformatics and biotechnology. Furthermore, association between genes and traits was analyzed. The main results are as follows:1. Pig ESTs were identified through standard BLAST analysis by using cDNA sequences of human homologous genes. Highly conserved pig ESTs were assembled to EST contigs. And primers selected from the contigs were used to isolate 34 novel porcine gene fragments from pig genomic DNA. The 34 novel porcine genes were as follows:(1) 20S proteasome alpha subunit genes PSMA1, PSMA2, PSMA3, PSMA5, PSMA6, PSMA7; beta subunit genes PSMB1, PSMB2, PSMB3, PSMB4, PSMB5, PSMB6, PSMB7, PSMB8, PSMB9, PSMB10; (2) 19S proteasome subunit genes PSMD7, PSMD8, PSMD9, PSMD10, PSMD11, PSMD12, PSMD13, PSMD14; (3) signalsome genes SGN1, SGN2, SGN3, SGN4, SGN5, SGN6, SGN7, SGN8; (4) ubiquitin specific protease genes USP6, USP10.2. Combining the in silico cloning and RT-PCR method, the entire coding region of the PSMB4, PSMB6, PSMB8, PSMB9, PSMB10 genes were obtained as well as the full length genomic DNA of PSMB4, PSMB6, PSMB8, PSMB9, PSMB10. The sequences were analyzed with species homologous genes and amino acid sequences were deduced to forecast functional domains.3. Expression profiles of PSMB4, PSMB6, PSMB8, PSMB10 genes of Tongcheng pigs were analysis by semi-quantitative RT-PCR assay and realtime-PCR. Except for PSMB8 gene unexpressing in heart and muscle, other genes are widely expressed in tissues of heart, liver, spleen, lung, kidney, muscle and fat. 4. 25 novel porcine genes were physical mapped by the RH panel. Results were showed as below: PSMA1 gene was located on SSC2, PSMA2 gene was located on SSC18, PSMA3 gene was located on SSC14, PSMA5 gene was located on SSC4, PSMA6 gene was located on SSC17, PSMB2 gene was located on SSC6, PSMB3 gene was located on SSC 12, PSMB4 gene was located on SSC4, PSMB6 gene was located on SSC12, PSMB7 gene was located on SSC1, PSMB8 gene was located on SSC7, PSMB9 gene was located on SSC7, PSMB10 gene was located on SSC6, PSMD8 gene was located on SSC6, PSMD9 gene was located on SSC14, PSMD10 gene was located on SSCX, PSMD12 gene was located on SSC12, PSMD13 gene was located on SSC2, PSMD14 gene was located on SSC15, SGN2 gene was located on SSC1, SGN4 gene was located on SSC8, SUN5 gene was located on SSC4, SGN6 gene was located on SSC3, USP6 gene was located on SSC12, USP10 gene was located on SSC6.5. Studies on DNA polymorphisms of PSMA1, PSMA6, PSMB4, PSMB6, PSMB8 and PSMB10 genes by PCR-RFLP combining DHLPC revealed ten polymorphic sites. Allele frequencies among different pig populations were determined.6. The all polymorphic sites were genotyped in the experimental pig population constructed under the cooperation of our lab and animal husbandry bureau of Tongcheng county (include 22 Landrace pigs, 23 Large White pigs, 58 Tongcheng pigs, 27 Landrace♂×(Large White×Tongcheng)♀pigs and 26 Large White♂×(Landrace×Tongcheng)♀pigs). Association of genotypes with phenotypes of production traits and immune traits were analyzed. The results showed that there are associations between the genotype of PSMA1, PSMA6, PSMB6, PSMB8, PSMB10 genes with some production traits and some immuno traits except PSMB4 gene.
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