Abstract:

【Objective】 The function of Calsarcin-1 was studied in the formation of skeletal muscle fiber and signaling pathway regulation through the screening for porcine Calsarcin-1 interacting proteins by yeast two-hybrid system. 【Method】 The pGBKT7-CS1 bait vector without toxicity or auto-activation was constructed with the Calsarcin-1 gene of Landrace pig, firstly. Then the mRNA was isolated from the skeletal muscle of the adult Landrace and purified ds-cDNA by SMART technology. Ds-cDNA was amplified by long distance PCR. At last, the Calsarcin-1 interaction proteins were screened through co-transformation with bait vector and ds-cDNA prey library. With the alignment of interacted gene in GenBank, the function of Calsarcin-1 was analyzed.【Result】 In this study, pGBKT7-CS1 bait expression vector was constructed. The ss-cDNAs of the porcine skeletal muscle with full length 3’ end were isolated and the ds-cDNAs were synthesized. Four proteins interacting with Calsarcin-1 were identified. The Calsarcin-1 showed the important function in maintaining the structure of Z line through interacting with ACTA1 and ACTN3, while involved to calcium ion regulation through CASQ1 and TNNT3 possibly.【Conclusion】 According to the result, it is presumed that the Calsarcin-1 affected the formation of Z-line and the calcium binding by combination with these identified proteins and participated in the calcium ion regulation and maintaining the cytoskeleton stabilization. By this way, the calcium ion concentration also affected other factors and signaling pathways related to the slow and fast muscle fiber formation and transformation.

Key words: porcine, skeletal muscle, Calsarcin-1 gene, yeast two-hybrid