Study On The Pocine CACNA1S Gene

The calcium channel of human skeletal muscle DHP-sensitive is a protein complex composed ofα₁,α₂,β,γandδsubunits, which form functional units and protein pores bound to the membrane. This protein complex controls the intra- and/or extracellular ion transportion and therefore, the function of the cell. The most important subunit of calcium channel that encoded by CACNA1S gene acts as a voltage-sensor, which possesses the selectivity of ion and provides the joint point for chemicals such as calcium antagonists and forms the transmembrane hydrophilic channel. Mutation in CACNA1S gene can cause hypokalemic periodic paralysis (HypoKPP) and malignant hyperthermia (MH) in human beings. The CACNA1S gene has been mapped to human chromosome 1q31 -32, but not reported in swine and other species. Following the fluorescence in situ hybridization (FISH), the radiation hybrid panel (RH) becomes a convenient and effective method for mapping genes. It could provide directive result and information for genes mapping. The primers were designed based on the published human CACNA1S 3' UTR sequence for ampilication and sequence of the porcine genome DNA, furthermore the alignment was done between human and porcine sequences, and the CACNA1S gene was mapped to the porcine chromosome 10p11-12 by using the IMpRH panel.Based on the published human CACNA1S gene, a sequence of 5380 bp for the CACNA1S gene was amplified, which include 2000 bp and 3380 bp for extron and intron repectively. The alignment result showed that there was 64.6%-97.4% identity between human and porcine extron sequences. The DNA sequence of 3045 bp from CACNA1S gene were examined by PCR-SSCP and sequencing, and 56 SNPs were detected, which include 25 SNPs for 1342 bp fragment in extron and 31 SNP's for 1703 bp fragment in intron. The polymorphism was detected in most of the primer pairs except for the 1S-47 and 1S-73, and 3 PCR-RFLP methods were developed based on the SNP's, all the pigs with the phenotype record were genotyped for those loci.The genetic effect of the loci on the carcass and meat quality trait was analyzed by the GLM procedure of SAS software, the results showed that: The locus 1S-46 has highly significant effect on water-holding capacity (P <0.01), significant effect on the L value of meat color, loin eye muscle area (0.01