STEAP4基因蛋白铁氧化还原酶活性分析  

Analysis of Iron Oxide Reductase Activity of STEAP4 Protein

作　　者：秦大妮[1] 季晨博[1] 张春梅[1] 史春梅[1] 朱冠忠[1] 郭锡熔[1] 

机构地区：[1]南京医科大学附属南京妇幼保健院儿科,南京210004

出　　处：《实用儿科临床杂志》2011年第7期485-486,496,共3页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(30772364);江苏省自然科学基金(BK2007230);江苏省医学领军人才项目(LJ200624);教育部2007年博士点基金联合资助课题(20070312001);江苏省研究生科研创新项目(CX09B_256Z)

摘　　要：目的分析肥胖相关基因STEAP4的铁氧化还原酶活性。方法体外温箱培养昆虫细胞株(Sf9),分别将pAcSec1-STEAP4转移载体和pAcSec1-vector转移载体与SapphireTM杆状病毒基因组混合后共转染Sf9昆虫细胞,通过G418药物筛选稳定表达STEAP4蛋白的细胞株,并抽提细胞总蛋白进一步应用Western blot技术验证其转染情况。应用分光光度计法检测铁氧化还原酶活性。应用SPSS11.5软件进行统计学分析。结果 Western blot技术证实STEAP4过表达细胞株构建成功;分光光度计法检测STEAP4铁氧化还原酶活性,STEAP4过表达组A值为5.182 5±0.230 4,空载组A值为1.615 0±0.822 6,STEAP4过表达组铁氧化还原酶活性明显高于空载组,差异有统计学意义(Pa<0.001)。结论 STEAP4基因蛋白具有铁氧化还原酶活性。Objective To analyze the iron oxide reductase activity of STEAP4 protein.Methods Insect cells（Sf9） were cultured in vitro,respectively,will pAcSec1-STEAP4 baculovirus transfer vector and pAcSec1-vector with the SapphireTM genomic mixture of Sf9 insect cells were transfected,and then selected by G418 and stable expression strain.After the extraction of protein,Western blot technique was applied to verify the transfection.Spectrophotometer was used to detect the iron oxide reductase activity of STEAP4.SPSS 11.5 software was used to analyze the data.Results Western blot technique results confirmed that the strain with over-expression of STEAP4 were constructed successfully,spectrophotometer assay STEAP4 iron oxide reductase activity,STEAP4 over-expression group absorbance value was 5.182 5±0.230 4 and vector group was 1.615 0±0.822 6,absorbance oxidoreductase activity in STEAP4 over-expression group was significantly higher than that in the other group,the differences were statistically significant（P0.001）.Conclusion STEAP4 gene has the activity of iron oxide reductase.

关 键 词：肥胖 STEAP4基因 铁氧化还原酶活性 

分 类 号：R589.2[医药卫生—内分泌]