【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。更多还原

【Abstract】 Construct pEGFP-NDPK B eukaryotic expression vector,forecast NDPK B protein 3D structure and activity 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 predict 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 inhibit the cellular growth,and has a positive regulatory role in cellular apoptosis.更多还原