NDPK B真核表达载体的构建及其对细胞生长活性的影响  

Construction of NDPK B eukaryotic expression vector and influence on cellular growth activity

作　　者：郭育培[1] 杨建新[1] 张学东[1] 王泽[1] 党源[1] 张晓东[1] 丁壮[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国兽医学报》2012年第7期1038-1042,1055,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972192);高校博士点基金资助项目(20110061110005)

摘　　要：将目的基因NDPK B连接到pEGFP-N1真核表达载体,转染Vero细胞,通过增强型绿色荧光蛋白(EGFP)和Western blotting分析,鉴定构建的pEGFP-NDPK B真核表达载体的正确性;利用生物信息学工具分析预测NDPKB蛋白的三维结构和可能存在的活性位点;MTT试验检测NDPK B对细胞生长活性的影响。结果显示,pEGFP-NDPK B真核表达载体构建成功,在Vero细胞中NDPK B和GFP蛋白能有效地融合表达。在线预测了NDPK B蛋白的三维结构和可能存在的活性位点。MTT试验结果表明,NDPK B对Vero细胞生长活性的抑制率为25.8%,对A549细胞生长活性的抑制率为22.7%。结果表明,NDPK B能够抑制细胞的生长,对细胞的凋亡具有正调节作用。Construct pEGFP-NDPK B eukaryotic expression vector,forecast NDPK B protein 3D structure and activi- ty sites,research the influence of NDPK B expression on cellular growth activity. Connected the NDPK B gene to pEGFP-N1 eukaryotic expression vector,transfected into Vero cells,through the enhanced green fluorescent protein （EGFP） and western blot identified the eukaryotic expression vector . Used the bioinformatics analysis tools to pre- dict NDPK B protein 3D structure and putative activity sites, MTT test detected the influence of NDPK B expression on cellular growth activity, pEGFP- NDPK B eukaryotic expression vector was constructed successfully, the NDPK B-GFP could express in Vero cells effectively, the 3D structure and activity sites of NDPK B had been predicted, the MTT results suggested the suppression ratio in Vero cells was 25.8% ,and 22.7% in A549 cells. NDPK B can inhib- it the cellular growth,and has a positive regulatory role in cellular apoptosis.

关 键 词：pEGFP-NDPK B真核表达载体 融合表达 三维结构 细胞生长活性 

分 类 号：S852.2[农业科学—基础兽医学]