中文摘要：

目的对比分析心肌CaV1.2钙通道片段GST-CT1融合蛋白与胞浆钙调蛋白（calmodulin,CaM）提取与纯化方法的异同。方法将pGEX-6p-3/CT1和pGEX-6p-3/CaM重组质粒转入E.coli BL21菌株,筛选得到转化成功的菌株并用IPTG诱导其表达,GS-4B beads分离纯化,SDS-PAGE凝胶电泳鉴定目的蛋白及其相对纯度,Pull down assay方法鉴定GST-CT1融合蛋白和CaM蛋白的生物学活性。结果采用超声破碎法,应用N-laurylsarcosine和Triton X-100处理后提取的GST-CT1融合蛋白浓度和纯度显著高于未用两者处理的对照组,而N-laurylsarcosine和Triton X-100对胞浆蛋白CaM的提取和纯化无明显影响。结论细胞膜通道蛋白GST-CT1的提取和纯化方法与胞浆蛋白Ca M存在明显差别。

英文摘要：

Objective To compare and analysis the purifying methods between CT1 fragment of L-type calcium channel CaV1.2 and calmodulin（CaM）.Methods pGEX-6p-3/CT1 and pGEX-6p-3/Ca M were transfected into E.coli BL21,and screening the bacterial strain with successful transfection,then stimulating the expression with IPTG,separating and purifying with GS-4B beads.SDS-PAGE was used to identify the purity of the target protein.The biological activity of the GST-CT1 and Ca M was detected by Pull down assay.Results N-laurylsarcosine and Triton X-100 could upregulate the concentration and purity of GST-CT1,but have no effect on CaM.Conclusion The purifying method for CT1 fragment of L-type calcium channel Ca_V 1.2 is obviously different from that for calmodulin.