【目的】维生素D3(vitamin D3,VD3)是一类重要的类固醇激素,参与调控动物体内多种生理活动,具有生物活性的VD3代谢物1,25(OH)2D3通过与维生素D受体(vitamin D receptor,VDR)结合发挥生物学功能。试验旨在克隆鹅VDR基因CDS序列并制备鹅VD...【目的】维生素D3(vitamin D3,VD3)是一类重要的类固醇激素,参与调控动物体内多种生理活动,具有生物活性的VD3代谢物1,25(OH)2D3通过与维生素D受体(vitamin D receptor,VDR)结合发挥生物学功能。试验旨在克隆鹅VDR基因CDS序列并制备鹅VDR多克隆抗体,为后续开展VDR介导VD3调控鹅体内多种生理活动的分子机制研究提供基础支撑。【方法】通过基因克隆方法获得鹅VDR基因CDS序列,并采用生物信息学方法对其编码蛋白进行结构和功能预测分析。通过基因合成和亚克隆方法构建鹅VDR基因重组表达质粒,转化大肠杆菌BL21(DE3)感受态细胞,经IPTG诱导表达和镍柱纯化后获得鹅VDR重组蛋白。以纯化的鹅VDR重组蛋白作为免疫原,制备兔抗鹅VDR多克隆抗体,采用间接ELISA法检测抗体效价,采用Western blotting检测抗体特异性。【结果】试验成功克隆了鹅VDR基因编码区序列,长度为1356 bp,编码451个氨基酸残基。该编码蛋白与鸡VDR蛋白的相似性高达91.4%,无信号肽和跨膜结构域,N-端含有核定位序列,且含有DNA结合域和配体结合域2个保守结构域,属于类固醇/甲状腺激素受体超家族成员。试验成功构建了pET-30a(+)-VDR重组质粒,并诱导表达鹅VDR重组蛋白。纯化后的重组蛋白纯度高达90%以上,且大小符合预期(52 ku)。试验制备了兔抗鹅VDR多克隆抗体,其抗体效价>1∶1093500;除肝脏组织外,该多克隆抗体在产蛋鹅肾脏、十二指肠、空肠和回肠组织中均能够识别天然VDR蛋白的2种亚型。【结论】试验成功克隆了鹅VDR基因CDS序列,并制备了兔抗鹅VDR多克隆抗体,该多克隆抗体特异性高,可用于后续鹅体内VDR介导的生物学功能研究。展开更多
以黄山毛峰为原料,采用实时荧光定量反转录聚合酶链式反应法和蛋白免疫印迹法研究其茶多酚提取物对肝药酶Cyp3a11及孕烷X受体(pregnane X receptor,PXR)的mRNA和蛋白表达影响,在此基础上采用瞬时共转染报告基因技术进一步验证是否通过PX...以黄山毛峰为原料,采用实时荧光定量反转录聚合酶链式反应法和蛋白免疫印迹法研究其茶多酚提取物对肝药酶Cyp3a11及孕烷X受体(pregnane X receptor,PXR)的mRNA和蛋白表达影响,在此基础上采用瞬时共转染报告基因技术进一步验证是否通过PXR实现对Cyp3a11表达调控。结果表明:黄山毛峰茶多酚提取物各剂量组(75、150、300 mg/(kg·d))均能显著调控小鼠肝Cyp3a11、PXR m RNA及其蛋白表达;报告基因实验结果显示,100、200、300μg/m L黄山毛峰茶多酚提取物经PXR通路使细胞色素P4503A4基因荧光素酶活力分别增加(3.86±0.05)、(4.82±0.72)、(5.38±0.11)倍。研究表明黄山毛峰茶多酚提取物可激活PXR通路调控肝药酶Cyp3a11的表达。展开更多
The variation in soil organic carbon density（SOCD） has been widely documented at various spatial and temporal scales. However, an accurate method for examining the attribution of explanatory factors for change in SO...The variation in soil organic carbon density（SOCD） has been widely documented at various spatial and temporal scales. However, an accurate method for examining the attribution of explanatory factors for change in SOCD is still lacking. This study aims to attribute and quantify the key climatic factors, anthropogenic activities, and soil properties associated with SOCD change in the native grasslands of Inner Mongolia, China, by comparing data between the 1960s and the 2010s. In 2007 and 2011, we resampled 142 soil profiles which were originally sampled during 1963–1964 in the native grasslands of Inner Mongolia. SOCD was determined in A horizon（eluvial horizon） of the soil. We selected the explanatory factors based on a random forest method, and explored the relationships between SOCD change and each of the explanatory factors using a linear mixed model. Our results indicated that the change in SOCD varied from the east to the west of Inner Mongolia, and SOCD was 18% lower in the 2010s than in the 1960s. The lower SOCD in the 2010s may primarily be attributed to the increasing in mean annual water surface evaporation, which explained approximately 10% and 50% of the total variation and explainable variation in the change in SOCD, respectively. The sand content of the soil is also a significant explanatory factor for the decrease in SOCD, which explained about 4% and 21% of the total variation and explainable variation in the change in SOCD, respectively. Furthermore, the collection of quantitative information on grazing frequency and duration may also help to improve our understanding of the anthropogenic factors that govern the change in SOCD.展开更多
文摘以黄山毛峰为原料,采用实时荧光定量反转录聚合酶链式反应法和蛋白免疫印迹法研究其茶多酚提取物对肝药酶Cyp3a11及孕烷X受体(pregnane X receptor,PXR)的mRNA和蛋白表达影响,在此基础上采用瞬时共转染报告基因技术进一步验证是否通过PXR实现对Cyp3a11表达调控。结果表明:黄山毛峰茶多酚提取物各剂量组(75、150、300 mg/(kg·d))均能显著调控小鼠肝Cyp3a11、PXR m RNA及其蛋白表达;报告基因实验结果显示,100、200、300μg/m L黄山毛峰茶多酚提取物经PXR通路使细胞色素P4503A4基因荧光素酶活力分别增加(3.86±0.05)、(4.82±0.72)、(5.38±0.11)倍。研究表明黄山毛峰茶多酚提取物可激活PXR通路调控肝药酶Cyp3a11的表达。
基金financially supported from the National Natural Science Foundation of China (41471093)the Key Technologies Research and Development Program of China (2012BAC19B04)+2 种基金the International Science and Technology Cooperation Project of China (2012DFA31290)the Fundamental Research Funds for Central Non-profit Scientific Institution (1610132016033,1610132016027)the Special Funding for the Modern Agricultural Technology System from the Chinese Ministry of Agriculture (CARS-35-11)
文摘The variation in soil organic carbon density（SOCD） has been widely documented at various spatial and temporal scales. However, an accurate method for examining the attribution of explanatory factors for change in SOCD is still lacking. This study aims to attribute and quantify the key climatic factors, anthropogenic activities, and soil properties associated with SOCD change in the native grasslands of Inner Mongolia, China, by comparing data between the 1960s and the 2010s. In 2007 and 2011, we resampled 142 soil profiles which were originally sampled during 1963–1964 in the native grasslands of Inner Mongolia. SOCD was determined in A horizon（eluvial horizon） of the soil. We selected the explanatory factors based on a random forest method, and explored the relationships between SOCD change and each of the explanatory factors using a linear mixed model. Our results indicated that the change in SOCD varied from the east to the west of Inner Mongolia, and SOCD was 18% lower in the 2010s than in the 1960s. The lower SOCD in the 2010s may primarily be attributed to the increasing in mean annual water surface evaporation, which explained approximately 10% and 50% of the total variation and explainable variation in the change in SOCD, respectively. The sand content of the soil is also a significant explanatory factor for the decrease in SOCD, which explained about 4% and 21% of the total variation and explainable variation in the change in SOCD, respectively. Furthermore, the collection of quantitative information on grazing frequency and duration may also help to improve our understanding of the anthropogenic factors that govern the change in SOCD.
