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转Bt基因大米与相应亲本大米差异蛋白质组学研究
引用本文:程娟献,夏晴,桑志红,毛劼,王心正,董方霆,何昆. 转Bt基因大米与相应亲本大米差异蛋白质组学研究[J]. 国际药学研究杂志, 2016, 0(4): 768-773. DOI: 10.13220/j.cnki.jipr.2016.04.035
作者姓名:程娟献  夏晴  桑志红  毛劼  王心正  董方霆  何昆
作者单位:国家生物医学分析中心, 北京,100850
基金项目:转基因生物新品种培育国家科技重大专项资助项目(2014ZX08011007);国家重大科学仪器设备开发专项资助项目(2012YQ180117)
摘    要:目的:利用差异蛋白质组学及生物信息学等技术方法,探讨外源Bt基因的导入对大米表达蛋白质组的影响,深化转基因大米在蛋白质组学方面的研究。方法在转基因大米“华恢1号”和亲本大米“明恢63”样品中提取大米总蛋白,通过二维聚丙烯酰胺凝胶电泳(2D-PAGE)实验方法,得到相应的蛋白质组2D-PAGE谱,再选择差异明显的蛋白质点进行质谱鉴定及生物信息学分析。结果对转Bt(cry1Ab/1Ac)基因大米和亲本大米2D-PAGE图谱的蛋白质点进行了对比匹配,识别出明显的差异蛋白质点28个,以亲本大米为参照,转Bt基因大米相对高表达的18个,相对低表达的10个;选择转基因大米凝胶上的差异蛋白质点进行了质谱鉴定和生物信息学检索,发现差异蛋白质主要参与能量代谢,蛋白质合成、氧化还原和应激响应等生物过程。结论转Bt基因“华恢1号”及其亲本大米“明恢63”表达的蛋白质组存在一定差异,但未发现这些差异蛋白质具有抗营养性和致敏性,也未发现新蛋白和毒蛋白的表达。

关 键 词:转基因大米  差异蛋白质组学  二维聚丙烯酰胺凝胶电泳  质谱鉴定

Differential proteomic analysis of transgenic Bt rice and its non-transgenic parental rice
CHENG Juan-xian;XIA Qing;SANG Zhi-hong;MAO Jie;WANG Xin-zheng;DONG Fang-ting;HE Kun. Differential proteomic analysis of transgenic Bt rice and its non-transgenic parental rice[J]. Foreign Medical Sciences(Section of Pharmarcy), 2016, 0(4): 768-773. DOI: 10.13220/j.cnki.jipr.2016.04.035
Authors:CHENG Juan-xian  XIA Qing  SANG Zhi-hong  MAO Jie  WANG Xin-zheng  DONG Fang-ting  HE Kun
Affiliation:CHENG Juan-xian;XIA Qing;SANG Zhi-hong;MAO Jie;WANG Xin-zheng;DONG Fang-ting;HE Kun;National Center of Biomedical Analysis;
Abstract:Objective Under the methodology of differential proteomics and bioinformatics,the impact of the exogenous gene on the expression of rice in the proteome is discussed,aiming to explore into the study of genetically modified rice in the proteomics. Methods The total protein was extracted from genetically modified rice Huahui No.1(HH1) and non-transgenic rice Minghui 63 (MH63),the method of two-dimensional gel electrophoresis was applied to generate corresponding proteome two-dimensional poly?acrylamide gel(2D-PAGE)electrophoresis spectrum;then,the mass spectrometry and bioinformatics analysis were conducted after the selection of protein spots with significant differences. Results The comparing and matching of protein spots between transgenic Bt (cry1Ab/1Ac)rice and non-transgenic rice 2D-PAGE profiles identified 28 protein spots with significant differences. With non-trans?genic rice as a reference,transgenic Bt rice held 18 relatively high and 10 relatively low expressions;mass spectrometry and bioinfor?matics retrieval were made on the different protein spots. It was found that the differentiated protein was mainly involved in energy me?tabolism,protein synthesis,redox stress response and other biological processes. Conclusion Differences exist between transgenic Bt HH1 and its parental rice MH63 on the expression of proteome;however,there are neither anti-nutritional and allergenic protein, nor new or toxic proteins among these differentiated proteins.
Keywords:transgenic rice  differential proteomics  two-dimensional polyacrylamide gel electrophoresis  mass spectrometry
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