酵母双杂交筛选猪Calsarcin-3基因相互作用蛋白的研究  

Yeast Two-hybrid Study of Pig Calsarcin-3 Interaction Proteins

作　　者：陈大雷[1,2] 朱文娟[1,3] 杨述林[1] 杨连玉[2] 崔文涛[1] 储明星[1] 马月辉[1] 李奎[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所农业部畜禽遗传资源与利用重点开放实验室,北京100193 [2]吉林农业大学动物科技学院,长春130118 [3]华中农业大学动物医学院,武汉430070

出　　处：《畜牧兽医学报》2009年第11期1594-1599,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金项目(30500359);国家863计划(2006AA10Z135;2008AA10Z143);973计划(2006CB102105);中国农业科学院北京畜牧兽医研究所创新团队基金资助

摘　　要：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Calsarcin-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。This work was conducted to study the regulation role of Calsarcin-3（CS3） and its interaction proteins on Calciumion signal transduction pathways and understand the molecular mechanisms 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 pGBKT7-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 capping 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.

关 键 词：猪 肌肉 Calsarcin-3 Z盘 酵母双杂交 

分 类 号：S828[农业科学—畜牧学] S813.3[农业科学—畜牧兽医]