NDPKB真核表达载体的构建及其对细胞生长活性的影响 |
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引用本文: | 郭育培,杨建新,张学东,王泽,党源,张晓东,丁壮.NDPKB真核表达载体的构建及其对细胞生长活性的影响[J].兽医大学学报,2012(7):1038-1042,1055. |
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作者姓名: | 郭育培 杨建新 张学东 王泽 党源 张晓东 丁壮 |
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作者单位: | 吉林大学畜牧兽医学院,吉林长春130062 |
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基金项目: | 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) |
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摘 要: | 将目的基因NDPKB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Westernblotting分析,鉴定构建的pEGFP—NDPKB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NDPKB对细胞生长活性的影响。结果显示,pEGFP-NDPKB真核表达载体构建成功,在Vero细胞中NDPKB和GFP蛋白能有效地融合表达。在线预测了NDPKB蛋白的三维结构和可能存在的活性位点。MTT试验结果表明,NDPKB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,NDPKB能够抑制细胞的生长,对细胞的凋亡具有正调节作用。
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关 键 词: | pEGFP—NDPK B真核表达栽体 融合表达 三维结构 细胞生长活性 |
Construction of NDPK B eukaryotic expression vector and influence on cellular growth activity |
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Authors: | GUO Yu-pei YANG Jian-xin ZHANG Xue-dong WANG Ze DANG Yuan ZHANG Xiao-dong DING Zhuang |
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Institution: | (College of Animal Science and Veterinary Medicine , J ilin University, Chang-chun 130062, China) |
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Abstract: | 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. |
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Keywords: | eukaryotic expression vector of pEGFP-NDPK B fusion expression 3D structure cellular growth activity |
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