[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR produ...[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR products of NfEBP1 with particular restriction sites and expression vectors, pET-28 and pET-16b were digested. Ligation, transformation and selection were performed to construct the recombinant plasmids pET-28-NfEBP1 and pET-16-NfEBP1. The recombinant plasmids were transformed into E. coli BL21 using heat -shock transformation. [Results] Recombinant plasmids pET-28-NfEBP1-1188 and pET-16-NfEBP1-1188 were constructed and transformed into expressional host cells, E. coli BL21, and validated by colony PCR, sequencing and double digestion. [Conclusion] Prokaryotic expression vectors of EBP1 gene from N. fordii were successfully constructed, which laid the foundation for characterization of the gene function.展开更多
目的考察青天葵Nervilia fordii(Hance)Schltr.氯仿、乙酸乙酯、正丁醇、乙醇提取物抗氧化活性与其总黄酮和总酚含有量的相关性。方法 Al Cl3-HAc-Na Ac显色法和福林-酚比色法分别测定青天葵4种提取物中总黄酮和总酚含有量,通过测定DPP...目的考察青天葵Nervilia fordii(Hance)Schltr.氯仿、乙酸乙酯、正丁醇、乙醇提取物抗氧化活性与其总黄酮和总酚含有量的相关性。方法 Al Cl3-HAc-Na Ac显色法和福林-酚比色法分别测定青天葵4种提取物中总黄酮和总酚含有量,通过测定DPPH自由基清除能力、羟自由基清除能力、ABTS自由基清除能力、还原能力评价提取物抗氧化活性,SPSS17.0软件分析抗氧化活性与2种成分含有量的相关性。结果青天葵4种提取物的抗氧化活性均呈量效关系,乙酸乙酯提取物最强,而乙醇提取物最弱。其与总黄酮含有量有一定相关性(0.604<r<0.638),与总酚含有量相关性显著(r>0.977)。结论青天葵乙酸乙酯提取物可用于天然抗氧化剂开发,总酚含有量是影响其抗氧化活性的主要因素。展开更多
采用RNeasy Plant Mini Kit法、RNAiso Plus法、RNAiso for Polysaccharide-rich PlantTissue法、CTAB法和SDS法,并结合4种抽提试剂组合,分别提取青天葵叶片总RNA。通过琼脂糖凝胶电泳、紫外检测及RT-PCR反应对提取效果进行了比较分析...采用RNeasy Plant Mini Kit法、RNAiso Plus法、RNAiso for Polysaccharide-rich PlantTissue法、CTAB法和SDS法,并结合4种抽提试剂组合,分别提取青天葵叶片总RNA。通过琼脂糖凝胶电泳、紫外检测及RT-PCR反应对提取效果进行了比较分析。结果表明:除RNAiso Plus法外,其它方法提取的总RNA均有一定程度的DNA污染。RNAiso Plus法组③(1/5体积KAC+4/5体积PCI)完整性最好,SDS法组④(1/5体积NaCl+4/5体积PCI)浓度最高,为372.4ng/μL。5种方法共20个处理组获得的RNA反转录后均能扩增出18SrRNA基因片段。该研究建立起了适用于青天葵叶片的总RNA提取方法,为青天葵的基因克隆、表达及转录组分析奠定了技术基础。展开更多
基金Supported by Research Fund of the Doctoral Program of Higher Education (200805720004)Scientific Research Foundation for Returned Scholars, Ministry of Education of China ([2009]1001)~~
文摘[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR products of NfEBP1 with particular restriction sites and expression vectors, pET-28 and pET-16b were digested. Ligation, transformation and selection were performed to construct the recombinant plasmids pET-28-NfEBP1 and pET-16-NfEBP1. The recombinant plasmids were transformed into E. coli BL21 using heat -shock transformation. [Results] Recombinant plasmids pET-28-NfEBP1-1188 and pET-16-NfEBP1-1188 were constructed and transformed into expressional host cells, E. coli BL21, and validated by colony PCR, sequencing and double digestion. [Conclusion] Prokaryotic expression vectors of EBP1 gene from N. fordii were successfully constructed, which laid the foundation for characterization of the gene function.
文摘目的考察青天葵Nervilia fordii(Hance)Schltr.氯仿、乙酸乙酯、正丁醇、乙醇提取物抗氧化活性与其总黄酮和总酚含有量的相关性。方法 Al Cl3-HAc-Na Ac显色法和福林-酚比色法分别测定青天葵4种提取物中总黄酮和总酚含有量,通过测定DPPH自由基清除能力、羟自由基清除能力、ABTS自由基清除能力、还原能力评价提取物抗氧化活性,SPSS17.0软件分析抗氧化活性与2种成分含有量的相关性。结果青天葵4种提取物的抗氧化活性均呈量效关系,乙酸乙酯提取物最强,而乙醇提取物最弱。其与总黄酮含有量有一定相关性(0.604<r<0.638),与总酚含有量相关性显著(r>0.977)。结论青天葵乙酸乙酯提取物可用于天然抗氧化剂开发,总酚含有量是影响其抗氧化活性的主要因素。
文摘采用RNeasy Plant Mini Kit法、RNAiso Plus法、RNAiso for Polysaccharide-rich PlantTissue法、CTAB法和SDS法,并结合4种抽提试剂组合,分别提取青天葵叶片总RNA。通过琼脂糖凝胶电泳、紫外检测及RT-PCR反应对提取效果进行了比较分析。结果表明:除RNAiso Plus法外,其它方法提取的总RNA均有一定程度的DNA污染。RNAiso Plus法组③(1/5体积KAC+4/5体积PCI)完整性最好,SDS法组④(1/5体积NaCl+4/5体积PCI)浓度最高,为372.4ng/μL。5种方法共20个处理组获得的RNA反转录后均能扩增出18SrRNA基因片段。该研究建立起了适用于青天葵叶片的总RNA提取方法,为青天葵的基因克隆、表达及转录组分析奠定了技术基础。
