ImageJ软件在重组质粒pET32a-CDK2中蛋白表达的应用  

Application of Image J software in analyzing protein expression of recombinant plasmid pET32a-CDK2

作　　者：陈炜烨[1] 刘冬冬[1] 徐建华[1] 陈丹娜[1] 何敏[1] 张战锋[1] 黄宪章[1] 

机构地区：[1]广东省中医院检验医学部,广东广州510120

出　　处：《中国热带医学》2014年第1期23-25,共3页China Tropical Medicine

基　　金：广东省科技计划项目(No.2010B060900069)

摘　　要：目的应用Image J软件探索细胞周期依赖性蛋白激酶2(CDK2)在重组质粒pET32a-CDK2中的蛋白表达条件。方法将重组质粒pET32a-CDK2转入表达菌株BL21(DE3),然后在不同时间点(0、1、2、3、4h)和不同浓度(0.5、1、2 mmol/L)异丙基-β-D-硫代半乳糖苷(IPTG)下诱导表达目的蛋白CDK2,对CDK2采用SDS-PAGE法进行蛋白电泳,并应用Image J软件对电泳条带进行灰度分析。结果 CDK2诱导表达量在不同时间点差异有统计学意义(P<0.001),其中0h与1h、2h、3h、4h诱导表达量的差异均有统计学意义(P=0.007,P<0.001,P<0.001,P<0.001);1h与3h和4h诱导表达量的差异均有统计学意义(P=0.001);而2h和1h、3h、4h诱导表达量的差异均无统计学意义(均有P>0.05);不同浓度IPTG下的CDK2诱导表达量差异无统计学意义(P=0.336,P=0.240,P=1.000)。结论根据Image J软件分析结果,采用0.5 mmol/L浓度IPTG 2h的条件,节约诱导时间和试剂用量。Objective To explore the protein expression conditions of the recombinant plasmid pET32a-CDK2 by the Image J software. Methods Transformed the pET32a-CDK2 plasmid into expression strains BL21（DE3）,then induced the CDK2 expression by different IPTG concentrations（0.5, 1, 2 mmol/L）. The CDK2 were detected by SDS-PAGE at different time points（0,1,2,3,4 h） and analyzed with Image J software. Results The expression of CDK2 has significant difference between different time points（0,1,2,3,4h）（P0.001）. there were statistical difference between time point 0h and the other time points（1,2,3,4h）（P = 0.007, P0.001, P0.001,P0.001）. There were also statistical difference between 1 h and 3 h and 4 h（P=0.001）. However, the expression of CDK2 were no statistical difference between 2h and 1h,3h and 4h（P=0.336, P=0.240,P=1.000）. There were no statistical difference in the CDK2 expression in different IPTG concentrations（P0.05）. Conclusions According to the analysis of Image J software, it suggested that the induced time and reagent level could be saved in the conditions of 2 hours with 0.5 mmol/L IPTG.

关 键 词：IMAGE J分析软件 细胞周期依赖性蛋白激酶2 原核表达 灰度分析 

分 类 号：R730.23[医药卫生—肿瘤]