以 p KB、pc DNA 3和 p CPA为原始质粒 ,构建 BL G启动子调控的组织型纤溶酶原激活剂 (t PA)乳腺组织特异性表达载体 p BTA。以 Sal I酶切质粒 p BTA,回收 5 .45 kb基因片段 BL G- t PA- b GHpoly A,通过显微注射 ,导入昆明小鼠受精卵...以 p KB、pc DNA 3和 p CPA为原始质粒 ,构建 BL G启动子调控的组织型纤溶酶原激活剂 (t PA)乳腺组织特异性表达载体 p BTA。以 Sal I酶切质粒 p BTA,回收 5 .45 kb基因片段 BL G- t PA- b GHpoly A,通过显微注射 ,导入昆明小鼠受精卵的雄原核内。共注 136 5枚 ,将存活的 10 5 3枚移植到 5 0只同期发情假孕母鼠的输卵管内。怀孕 2 4只 ,共产仔79只 ,上述仔鼠通过 PCR检测 ,结果 19只阳性。Southern blotting检测上述 19份 PCR阳性小鼠基因组 ,其中 4只为整合阳性 ,说明已获得了乳球蛋白启动子调控下的 t PA转基因小鼠 ,为 t PA在小鼠体内表达研究提供了必要条件。展开更多
目的:通过分析白花鬼针草不同组织中候选糖基转移酶基因的表达规律,初步筛选可能参与聚多炔糖苷生物合成途径的糖基转移酶,为白花鬼针草的糖基转移酶基因的研究提供数据支持。方法:本研究利用白花鬼针草转录组数据库中的DN19630等19个...目的:通过分析白花鬼针草不同组织中候选糖基转移酶基因的表达规律,初步筛选可能参与聚多炔糖苷生物合成途径的糖基转移酶,为白花鬼针草的糖基转移酶基因的研究提供数据支持。方法:本研究利用白花鬼针草转录组数据库中的DN19630等19个候选糖基转移酶基因的DNA序列,设计特异性引物。通过qRT-PCR技术检测候选糖基转移酶基因在白花鬼针草叶、茎、果、花、根中相对表达量。结果:通过对DN19630等19个候选糖基转移酶基因PCR产物扩增情况、扩增曲线图、熔解曲线图及相对表达量进行综合分析,发现DN19630、DN18211、DN12914、DN6852、DN5000g2、DN38310、DN14409g1、DN5293、DN34556、DN14409g2、DN15831这11个候选糖基转移酶基因在叶中表达量相对较高,在根中表达量最低;而DN3608在茎中表达量最高,根中表达量较低;DN21732、DN14031在花中表达量较高,在根中表达量最低。结论:DN19630等14个糖基转移酶可能参与聚多炔糖苷类化合物的生物合成。Objective: By analyzing the expression patterns of candidate glycosyltransferase genes in different tissues of Bidens pilosa var. radiata, we initially screened the glycosyltransferases that may be involved in the biosynthetic pathway of polyacetylene glycosides, providing data support for the study of glycosyltransferase genes in B. pilosa var. radiata. Methods: This study used the DNA sequences of 19 candidate glycosyltransferase genes such as DN19630 from the transcriptome database of B. pilosa var. radiata to design specific primers. The relative expression levels of the selected glycosyltransferase genes in the leaves, stems, fruits, flowers, and roots of B. pilosa var. radiata were determined through qRT-PCR technology. Results: Through comprehensive analysis of the PCR product amplification status, amplification curves, melting curves, and relative expression levels of 19 candidate glycosyltransferase genes including DN19630, it was found that the expression levels of 11 candidate glycosyltransferase genes, namely DN19630, DN18211, DN12914, DN6852, DN5000g2, DN38310, DN14409g1, DN5293, DN34556, DN14409g2, and DN15831, were relatively high in leaves and lowest in roots. In contrast, DN3608 exhibited the highest expression level in stems and a lower expression level in roots. Additionally, DN21732 and DN14031 showed higher expression levels in flowers and the lowest expression levels in roots. Conclusion: A total of 14 glycosyltransferases, including DN19630, are potentially involved in the biosynthesis of polyene glycosides.展开更多
文摘目的:通过分析白花鬼针草不同组织中候选糖基转移酶基因的表达规律,初步筛选可能参与聚多炔糖苷生物合成途径的糖基转移酶,为白花鬼针草的糖基转移酶基因的研究提供数据支持。方法:本研究利用白花鬼针草转录组数据库中的DN19630等19个候选糖基转移酶基因的DNA序列,设计特异性引物。通过qRT-PCR技术检测候选糖基转移酶基因在白花鬼针草叶、茎、果、花、根中相对表达量。结果:通过对DN19630等19个候选糖基转移酶基因PCR产物扩增情况、扩增曲线图、熔解曲线图及相对表达量进行综合分析,发现DN19630、DN18211、DN12914、DN6852、DN5000g2、DN38310、DN14409g1、DN5293、DN34556、DN14409g2、DN15831这11个候选糖基转移酶基因在叶中表达量相对较高,在根中表达量最低;而DN3608在茎中表达量最高,根中表达量较低;DN21732、DN14031在花中表达量较高,在根中表达量最低。结论:DN19630等14个糖基转移酶可能参与聚多炔糖苷类化合物的生物合成。Objective: By analyzing the expression patterns of candidate glycosyltransferase genes in different tissues of Bidens pilosa var. radiata, we initially screened the glycosyltransferases that may be involved in the biosynthetic pathway of polyacetylene glycosides, providing data support for the study of glycosyltransferase genes in B. pilosa var. radiata. Methods: This study used the DNA sequences of 19 candidate glycosyltransferase genes such as DN19630 from the transcriptome database of B. pilosa var. radiata to design specific primers. The relative expression levels of the selected glycosyltransferase genes in the leaves, stems, fruits, flowers, and roots of B. pilosa var. radiata were determined through qRT-PCR technology. Results: Through comprehensive analysis of the PCR product amplification status, amplification curves, melting curves, and relative expression levels of 19 candidate glycosyltransferase genes including DN19630, it was found that the expression levels of 11 candidate glycosyltransferase genes, namely DN19630, DN18211, DN12914, DN6852, DN5000g2, DN38310, DN14409g1, DN5293, DN34556, DN14409g2, and DN15831, were relatively high in leaves and lowest in roots. In contrast, DN3608 exhibited the highest expression level in stems and a lower expression level in roots. Additionally, DN21732 and DN14031 showed higher expression levels in flowers and the lowest expression levels in roots. Conclusion: A total of 14 glycosyltransferases, including DN19630, are potentially involved in the biosynthesis of polyene glycosides.
