目的:建立可用于甘草HMGR,SQS1,β-AS合酶基因CNVs测定的稳定可靠的检测体系。方法:利用Real timePCR方法对甘草HMGR,SQS1,β-AS合酶基因CNVs进行测定。结果:在HMGR,SQS1,β-AS合酶基因以及内标基因lectin的定量检测实验中,其定量反应...目的:建立可用于甘草HMGR,SQS1,β-AS合酶基因CNVs测定的稳定可靠的检测体系。方法:利用Real timePCR方法对甘草HMGR,SQS1,β-AS合酶基因CNVs进行测定。结果:在HMGR,SQS1,β-AS合酶基因以及内标基因lectin的定量检测实验中,其定量反应的Ct值分别在25.82～25.88,29.01～29.08,15.52～15.56,19.06～19.08变化,SD分别是0.033,0.032,0.024,0.011,其CV分别是0.12%,0.22%,0.16%,0.06%。结论:所建立的Real time PCR体系重复性好,检测系统工作稳定可靠,可用于甘草HMGR,SQS1,β-AS基因的CNVs筛选。展开更多
目的:对甘草道地性形成机制进行探索。方法:在GenBank中检索β-香树酯醇合成酶(β-AS)基因,进行序列比对并找出序列保守区,用primer premier 5.0软件设计引物,对3个不同产地甘草材料的β-AS基因进行扩增、测序,用MegAlign软件进行序列...目的:对甘草道地性形成机制进行探索。方法:在GenBank中检索β-香树酯醇合成酶(β-AS)基因,进行序列比对并找出序列保守区,用primer premier 5.0软件设计引物,对3个不同产地甘草材料的β-AS基因进行扩增、测序,用MegAlign软件进行序列分析并构建系统树。在表达差异实验中,对来自3个产地的9份甘草材料进行总RNA提取,逆转录得到cDNA,以甘草18S基因为内参,PCR扩增后进行相对表达量分析。结果:在序列多态性实验中,9个样品的β-AS序列经比对分析共发现108处变异位点,内含子变异位点数高于外显子变异位点数两倍。在表达差异实验中内蒙组、甘肃组和宁夏组的甘草β-AS基因平均相对表达量有显著性差异。结论:不同产地甘草β-AS基因多态性的差异以及表达量的差异,可能是导致甘草道地性形成的原因之一。展开更多
Glycyrrhiza uralensis is frequently used in traditional Chinese medicine. This plant contains a large amount of effective constituents, including triterpenoids and flavonoids. Among them, glycyrrhizin is believed to b...Glycyrrhiza uralensis is frequently used in traditional Chinese medicine. This plant contains a large amount of effective constituents, including triterpenoids and flavonoids. Among them, glycyrrhizin is believed to be the marker compound to evaluate the quality of G. uralensis based on Chinese Pharmacopoeia. Many studies showed that glycyrrhizin possesses various pharmacological activities, such as antibacterial, antiviral, antitumor, anti-inflammatory, and immune-stimulating activities. In this paper, we summarized the cloning, characterization, expression, and polymorphism analysis of several functional genes involved in glycyrrhizin biosynthesis in G. uralensis.展开更多
Objective:To study the influence of over-expression of three functional genes involved in GC biosynthetic pathway,GuHMGR,GuSQS1,and GuBAS on GC production.Methods:Three plant expression vectors were constructed and tr...Objective:To study the influence of over-expression of three functional genes involved in GC biosynthetic pathway,GuHMGR,GuSQS1,and GuBAS on GC production.Methods:Three plant expression vectors were constructed and transformed into Agrobacterium tumefaciens EHA105,which were used to infect Glycyrrhiza uralensis hypocotyls explants.After induction,selection,differentiation,culture,and transplantation,12,15,and 5 regenerated plants over-expressing GuHMGR,GuSQS1,and GuBAS,were obtained,respectively.Results:RT-PCR analysis showed these transgenic regenerated G.uralensis plants had 2-6 copies of GuHMGR,GuSQS1,or GuBAS.HPLC analysis showed the contents of 18α-and 18β-GC in all transgenic regenerated samples were both higher than that in the blank control.With the increase of copy numbers of GuHMGR,GuSQS1,and GuBAS,the contents of 18α-and 18β-GC were both increased in most samples.The highest 18α-and 18β-GC contents in transgenic regenerated plants were about 3.05 times and 2.80 times higher than that in the blank control,respectively.Conclusion:Over-expression of the GuHMGR,GuSQS1,and GuBAS genes enhance the accumulation of 18α-and 18β-GC in the roots and rhizomes of G.uralensis.We hope this work can lay a foundation for the molecular breeding research of G.uralensis and improving the quality of the roots and rhizomes of G.uralensis cultivars.展开更多
系统研究、改进和优化了甘薯淀粉加工废水中多酚氧化酶、β-淀粉酶、储藏蛋白、小分子有机物等生化成分的分离回收方法和工艺.通过等电点(p H 3.8)沉淀分离获得多酚氧化酶粗酶制剂;超滤浓缩和絮凝沉淀回收获得β-淀粉酶的粗酶制剂;纳滤...系统研究、改进和优化了甘薯淀粉加工废水中多酚氧化酶、β-淀粉酶、储藏蛋白、小分子有机物等生化成分的分离回收方法和工艺.通过等电点(p H 3.8)沉淀分离获得多酚氧化酶粗酶制剂;超滤浓缩和絮凝沉淀回收获得β-淀粉酶的粗酶制剂;纳滤膜浓缩回收小分子物质,2种粗酶制剂经体积分数50%乙醇沉淀可获得纯化的PPO和β-淀粉酶,上清液浓缩干燥获得甘薯储藏蛋白.该工艺可从每升甘薯淀粉加工废水中回收多酚氧化酶3.2 g(酶活性1.2×10~5U/g,回收率38.7%),β-淀粉酶1.2 g(酶活性3.4×10~7U/g,回收率97.5%),小分子物质7.2 g,储藏蛋白13.6 g,为提高甘薯的开发利用价值、解决甘薯淀粉生产业对环境的污染提供了新途径.展开更多
β-carotene with double fluorescence characteristics and large third-order optical nonlinearities,which is dissolved in the carbon disulfide(CS2) as the core medium of a liquid core optical fiber(LCOF),is applied in t...β-carotene with double fluorescence characteristics and large third-order optical nonlinearities,which is dissolved in the carbon disulfide(CS2) as the core medium of a liquid core optical fiber(LCOF),is applied in the study of the CS2 stimulated Raman scattering(SRS). The results of this study show that when the concentrations of solution are more than 3.72×10-7 mol/L,the amplified spontaneous emis-sion(ASE) of β-carotene is the main factor influencing the threshold and intensity of Stokes lines; when the concentrations of solution are lower than 3.72×10-7 mol/L,the ASE disappears and the fluorescence plays the key role:The high-order Stokes lines may be observed at very low input-laser power,and the Stokes thresholds decrease as the solution concentration increases. The result may be widely used in the study of broadband stimulated radiation laser and seeding laser.展开更多
By recombinant DNA technology, the N-terminal of the β-protein encoding region of plasmid pUBHO is fused with the structure gene of α-amylase from Bacillus licheniformis. This gene fusion is called βAmy. It is able...By recombinant DNA technology, the N-terminal of the β-protein encoding region of plasmid pUBHO is fused with the structure gene of α-amylase from Bacillus licheniformis. This gene fusion is called βAmy. It is able to transcribe and translate in phase. Protein fusion can be secreted into the medium mediated by β-signal peptide. The efficiency of secretion is about 10% of the synthesized pre-α-amylase. By comparing the secretion capacities and analysing the restriction sites on β-Amy genes and the molecular weights of the mature α-amylase secreted by B. subtilis harbouring different plasmids, it is indicated in vivo that the recognition and cleavage sequence for signal peptidase I of B. subtilis is Ala-Ala-Ala Ala. The results also indicate that the secretion of the α-amylase in B. subtilis is in accordance with the post-translational transportation mode.展开更多
文摘目的:建立可用于甘草HMGR,SQS1,β-AS合酶基因CNVs测定的稳定可靠的检测体系。方法:利用Real timePCR方法对甘草HMGR,SQS1,β-AS合酶基因CNVs进行测定。结果:在HMGR,SQS1,β-AS合酶基因以及内标基因lectin的定量检测实验中,其定量反应的Ct值分别在25.82～25.88,29.01～29.08,15.52～15.56,19.06～19.08变化,SD分别是0.033,0.032,0.024,0.011,其CV分别是0.12%,0.22%,0.16%,0.06%。结论:所建立的Real time PCR体系重复性好,检测系统工作稳定可靠,可用于甘草HMGR,SQS1,β-AS基因的CNVs筛选。
文摘目的:对甘草道地性形成机制进行探索。方法:在GenBank中检索β-香树酯醇合成酶(β-AS)基因,进行序列比对并找出序列保守区,用primer premier 5.0软件设计引物,对3个不同产地甘草材料的β-AS基因进行扩增、测序,用MegAlign软件进行序列分析并构建系统树。在表达差异实验中,对来自3个产地的9份甘草材料进行总RNA提取,逆转录得到cDNA,以甘草18S基因为内参,PCR扩增后进行相对表达量分析。结果:在序列多态性实验中,9个样品的β-AS序列经比对分析共发现108处变异位点,内含子变异位点数高于外显子变异位点数两倍。在表达差异实验中内蒙组、甘肃组和宁夏组的甘草β-AS基因平均相对表达量有显著性差异。结论:不同产地甘草β-AS基因多态性的差异以及表达量的差异,可能是导致甘草道地性形成的原因之一。
基金Beijing Project for Young Talents(YETP0819)National Science Foundation of China(81503181)
文摘Glycyrrhiza uralensis is frequently used in traditional Chinese medicine. This plant contains a large amount of effective constituents, including triterpenoids and flavonoids. Among them, glycyrrhizin is believed to be the marker compound to evaluate the quality of G. uralensis based on Chinese Pharmacopoeia. Many studies showed that glycyrrhizin possesses various pharmacological activities, such as antibacterial, antiviral, antitumor, anti-inflammatory, and immune-stimulating activities. In this paper, we summarized the cloning, characterization, expression, and polymorphism analysis of several functional genes involved in glycyrrhizin biosynthesis in G. uralensis.
文摘Objective:To study the influence of over-expression of three functional genes involved in GC biosynthetic pathway,GuHMGR,GuSQS1,and GuBAS on GC production.Methods:Three plant expression vectors were constructed and transformed into Agrobacterium tumefaciens EHA105,which were used to infect Glycyrrhiza uralensis hypocotyls explants.After induction,selection,differentiation,culture,and transplantation,12,15,and 5 regenerated plants over-expressing GuHMGR,GuSQS1,and GuBAS,were obtained,respectively.Results:RT-PCR analysis showed these transgenic regenerated G.uralensis plants had 2-6 copies of GuHMGR,GuSQS1,or GuBAS.HPLC analysis showed the contents of 18α-and 18β-GC in all transgenic regenerated samples were both higher than that in the blank control.With the increase of copy numbers of GuHMGR,GuSQS1,and GuBAS,the contents of 18α-and 18β-GC were both increased in most samples.The highest 18α-and 18β-GC contents in transgenic regenerated plants were about 3.05 times and 2.80 times higher than that in the blank control,respectively.Conclusion:Over-expression of the GuHMGR,GuSQS1,and GuBAS genes enhance the accumulation of 18α-and 18β-GC in the roots and rhizomes of G.uralensis.We hope this work can lay a foundation for the molecular breeding research of G.uralensis and improving the quality of the roots and rhizomes of G.uralensis cultivars.
基金Supported by the National Natural Science Foundation of China(Grant No.10774057)
文摘β-carotene with double fluorescence characteristics and large third-order optical nonlinearities,which is dissolved in the carbon disulfide(CS2) as the core medium of a liquid core optical fiber(LCOF),is applied in the study of the CS2 stimulated Raman scattering(SRS). The results of this study show that when the concentrations of solution are more than 3.72×10-7 mol/L,the amplified spontaneous emis-sion(ASE) of β-carotene is the main factor influencing the threshold and intensity of Stokes lines; when the concentrations of solution are lower than 3.72×10-7 mol/L,the ASE disappears and the fluorescence plays the key role:The high-order Stokes lines may be observed at very low input-laser power,and the Stokes thresholds decrease as the solution concentration increases. The result may be widely used in the study of broadband stimulated radiation laser and seeding laser.
基金Basic research of "863" High Technology Program, supported by the National Natural Science Foundation of China.
文摘By recombinant DNA technology, the N-terminal of the β-protein encoding region of plasmid pUBHO is fused with the structure gene of α-amylase from Bacillus licheniformis. This gene fusion is called βAmy. It is able to transcribe and translate in phase. Protein fusion can be secreted into the medium mediated by β-signal peptide. The efficiency of secretion is about 10% of the synthesized pre-α-amylase. By comparing the secretion capacities and analysing the restriction sites on β-Amy genes and the molecular weights of the mature α-amylase secreted by B. subtilis harbouring different plasmids, it is indicated in vivo that the recognition and cleavage sequence for signal peptidase I of B. subtilis is Ala-Ala-Ala Ala. The results also indicate that the secretion of the α-amylase in B. subtilis is in accordance with the post-translational transportation mode.
