CAJ | 학술논문

旨在研究猪Calsarcin-3(CS3)基因及其互作蛋白对肌细胞钙离子的调控作用,了解快慢肌纤维分化和转化的分子机理.从猪的骨骼肌组织样中提取mRNA,合成并纯化双链cDNA;然后用长白猪的Calsarcin-3基因构建既无自激活活性又无毒性的pGBKT7-CS3诱饵载体;最后通过酵母双杂交系统的杂交方法筛选与Calsarein-3相互作用的蛋白,在GenBank中对相互作用基因进行比对分析,分析Calsarcin-3在骨骼肌中的功能.通过筛选,共筛选出5个与Calsarcin-3基因互作的蛋白,其中,重要的蛋白有帽蛋白Zβ(CAPZB)、肌联蛋白(TTN)和α-肌动蛋白1(ACTA1).结果提示,CAPZB可能与Calsarcin-3共同作用参与尖锐端肌动蛋白丝戴帽;ACTA1可能与Calsar-cin-3共同作用作为一种收缩蛋白在骨骼肌的活动中发挥重要作用;TTN能与Calsarcin-3结合参与维持Z线结构的稳定.
지재연구저Calsarcin-3(CS3)기인급기호작단백대기세포개리자적조공작용,료해쾌만기섬유분화화전화적분자궤리.종저적골격기조직양중제취mRNA,합성병순화쌍련cDNA;연후용장백저적Calsarcin-3기인구건기무자격활활성우무독성적pGBKT7-CS3유이재체;최후통과효모쌍잡교계통적잡교방법사선여Calsarein-3상호작용적단백,재GenBank중대상호작용기인진행비대분석,분석Calsarcin-3재골격기중적공능.통과사선,공사선출5개여Calsarcin-3기인호작적단백,기중,중요적단백유모단백Zβ(CAPZB)、기련단백(TTN)화α-기동단백1(ACTA1).결과제시,CAPZB가능여Calsarcin-3공동작용삼여첨예단기동단백사대모;ACTA1가능여Calsar-cin-3공동작용작위일충수축단백재골격기적활동중발휘중요작용;TTN능여Calsarcin-3결합삼여유지Z선결구적은정.
This work was conducted to study the regulation role of Calsarcin-3 (CS3) and its inter-action proteins on Calciumion signal transduction pathways and understand the molecular mecha-nisms of skeletal muscle fiber differentiation and development. The mRNA was extracted from the skeletal muscle of the adult Landrace and the purified ds-cDNA was obtained. The pGBKTT-CS3 bait vector without toxicity and auto-activation was constructed with the Calsarcin-3 gene of Landrace pig. ds-cDNA was amplified by long distance PCR. Finally, the Calsarcin-3 interaction proteins were screened through hybridization of bait vector with ds-cDNA prey library. Through the alignment of interaction genes in GenBank, the function of Calsarcin-3 was analyzed. In this study, five proteins interacting with CS3 were identified. These results indicated that the β cap-ping protein muscle Z-line(CAPZB) may regulate the growth of the actin filament by capping the barbed end of growing actin filaments with Calsarcin-3. Actin, alpha 1 (ACTA1) as a contractile protein may play a significant role in regulating the skeletal muscle activity with Calsarcin-3. Titin (TTN) may be involed in the structure of the Z-disk by binding with Calsarcin-3.