[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene ...[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene in cotton bollworm. [Method] Total RNA was extracted from midgut of cotton bollworm. The full length cDNA of Habeta5 gene was cloned by using rapid amplification of cDNA ends (RACE) technology, then sequence analysis was carried out. [Result] The full length cDNA sequence was successfully cloned and isolated, named as Habeta5. It was 947 bp in length, contained an ORF (843 bp) and encoded 280 amino acid residues, with the predicted mass of 30.87 kD and isoelectric point(pI) of 9.60. In the deduced amino acid sequence, a proteasome β5 subunit domain lies between 74th to 261st amino acid residues. It has more than 62% identity to other insects such as Drosophila melanogaster. The proteasome β5 subunit conservative regions were very similar with each other. Molecular evolution by Neighbor Joining method indicated that Habeta5 was homologous with other proteasome β5 subunit of species. [Conclusion] Sequence alignment shows that the cloned fragment is a proteasome β5 subunit gene (GenBank accession number: FJ358434).展开更多
为了优化大孔吸附树脂分离纯化茼蒿总黄酮(TFC)的工艺条件,试验以TFC吸附量和回收率为指标,考察5种大孔吸附树脂静态、动态的吸附及解吸附性能。优选影响因素——上样浓度,上样p H值,洗脱浓度和洗脱流速4种试验条件,测定大孔树脂的泄露...为了优化大孔吸附树脂分离纯化茼蒿总黄酮(TFC)的工艺条件,试验以TFC吸附量和回收率为指标,考察5种大孔吸附树脂静态、动态的吸附及解吸附性能。优选影响因素——上样浓度,上样p H值,洗脱浓度和洗脱流速4种试验条件,测定大孔树脂的泄露曲线和洗脱曲线。研究发现,D-101大孔树脂对TFC的吸附纯化显示出最佳的性能。泄露曲线显示:上样体积泄漏点60 m L,饱和点210 m L。洗脱曲线显示:体积分数95%乙醇溶液80 m L洗脱效果最好。响应面试验方差分析显示:上样p H对TFC回收率影响差异达到显著水平,而上样浓度、洗脱浓度和洗脱速度3个因素对TFC回收率影响差异不显著。通过回归方程模拟预测最优纯化条件组合——上样浓度0.86mg/m L,p H=6,洗脱浓度74.27%,洗脱流速2.23 m L/min,TFC回收率由65.19%提升至87.36%。D-101大孔树脂对于TFC的分离纯化性能较好,适用于工业化推广。展开更多
基金Supported by National Basic Research Program of China(2005CB121005)~~
文摘[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene in cotton bollworm. [Method] Total RNA was extracted from midgut of cotton bollworm. The full length cDNA of Habeta5 gene was cloned by using rapid amplification of cDNA ends (RACE) technology, then sequence analysis was carried out. [Result] The full length cDNA sequence was successfully cloned and isolated, named as Habeta5. It was 947 bp in length, contained an ORF (843 bp) and encoded 280 amino acid residues, with the predicted mass of 30.87 kD and isoelectric point(pI) of 9.60. In the deduced amino acid sequence, a proteasome β5 subunit domain lies between 74th to 261st amino acid residues. It has more than 62% identity to other insects such as Drosophila melanogaster. The proteasome β5 subunit conservative regions were very similar with each other. Molecular evolution by Neighbor Joining method indicated that Habeta5 was homologous with other proteasome β5 subunit of species. [Conclusion] Sequence alignment shows that the cloned fragment is a proteasome β5 subunit gene (GenBank accession number: FJ358434).
文摘为了优化大孔吸附树脂分离纯化茼蒿总黄酮(TFC)的工艺条件,试验以TFC吸附量和回收率为指标,考察5种大孔吸附树脂静态、动态的吸附及解吸附性能。优选影响因素——上样浓度,上样p H值,洗脱浓度和洗脱流速4种试验条件,测定大孔树脂的泄露曲线和洗脱曲线。研究发现,D-101大孔树脂对TFC的吸附纯化显示出最佳的性能。泄露曲线显示:上样体积泄漏点60 m L,饱和点210 m L。洗脱曲线显示:体积分数95%乙醇溶液80 m L洗脱效果最好。响应面试验方差分析显示:上样p H对TFC回收率影响差异达到显著水平,而上样浓度、洗脱浓度和洗脱速度3个因素对TFC回收率影响差异不显著。通过回归方程模拟预测最优纯化条件组合——上样浓度0.86mg/m L,p H=6,洗脱浓度74.27%,洗脱流速2.23 m L/min,TFC回收率由65.19%提升至87.36%。D-101大孔树脂对于TFC的分离纯化性能较好,适用于工业化推广。
