目的:构建草原兔尾鼠卵透明带3 DNA疫苗pVAX1-sig-LTB-lZP3-C3d3进行小鼠的黏膜免疫,增强该疫苗的免疫不育效果。方法:将两种佐剂分子大肠杆菌不耐热肠毒素B亚基和C3d基因通过酶切鉴定,构建重组质粒pVAX1-sig-LTB-lZP3-C3d3,采用RT-PCR...目的:构建草原兔尾鼠卵透明带3 DNA疫苗pVAX1-sig-LTB-lZP3-C3d3进行小鼠的黏膜免疫,增强该疫苗的免疫不育效果。方法:将两种佐剂分子大肠杆菌不耐热肠毒素B亚基和C3d基因通过酶切鉴定,构建重组质粒pVAX1-sig-LTB-lZP3-C3d3,采用RT-PCR和W estern b lot技术,检测其在mRNA和蛋白水平的表达,并通过肌肉注射、滴鼻和灌服3种途径免疫雌性C57BL/6小鼠,通过ELISA检测抗体水平及分型。结果:酶切鉴定,RT-PCR和W estern b lot结果表明重组质粒构建正确并可在mRNA和蛋白水平的表达,ELISA结果表明以重组质粒pVAX1-sig-LTB-lZP3-C3d3免疫诱导的特异性IgG、IgA的水平明显高于对照组(P<0.01)。抗生育实验表明该疫苗免疫的小鼠平均生仔数与对照组相比差异极显著(P<0.01)。结论:构建的草原兔尾鼠卵透明带3DNA疫苗重组质粒pVAX1-sig-LTB-lZP3-C3d3可高效地激发小鼠特异性免疫应答,提高抗生育的效果。展开更多
A novel porous silicon based optical Fabry-Perot cavity structure is prepared as a label-free immunosensor platform for detecting antigen-antibody.The lagurus zona pelluciad 3(LZP3) and the specificity of the polyclon...A novel porous silicon based optical Fabry-Perot cavity structure is prepared as a label-free immunosensor platform for detecting antigen-antibody.The lagurus zona pelluciad 3(LZP3) and the specificity of the polyclonal anti-LZP3 antibodies are employed in our laboratory as the target and the probe,respectively.Firstly,the antibodies are immobilized to the porous silicon optical cavity using silanization and glutaraldehyde(GA) chemistry.And then,after the antigen-antibody reaction,it is monitored that the red shift of the reflection spectrum of the immunosensor increases with the antigen concentration.This research also plays a potential role for the extensive applications in immunoassay.展开更多
文摘目的筛选不同生长时期的油莎草地上茎的内参基因,为分析其基因的表达提供支持.方法以生长40 d的油莎草幼苗10:00,12:00,14:00,16:00,18:00,20:00六个时间点的地上茎为材料;以及生长天数为20,30,40,50,55,65,75,85,95,105,115,125,135,145 d的油莎草地上茎为材料,采用实时荧光定量PCR(Quantitative real-time PCR,RT-q PCR)技术,分析其18S r RNA,GAPDH-两个候选基因.结论筛选出18S r RNA作为分析油莎草地上茎不同时期基因表达量的内参基因.
文摘目的:构建草原兔尾鼠卵透明带3 DNA疫苗pVAX1-sig-LTB-lZP3-C3d3进行小鼠的黏膜免疫,增强该疫苗的免疫不育效果。方法:将两种佐剂分子大肠杆菌不耐热肠毒素B亚基和C3d基因通过酶切鉴定,构建重组质粒pVAX1-sig-LTB-lZP3-C3d3,采用RT-PCR和W estern b lot技术,检测其在mRNA和蛋白水平的表达,并通过肌肉注射、滴鼻和灌服3种途径免疫雌性C57BL/6小鼠,通过ELISA检测抗体水平及分型。结果:酶切鉴定,RT-PCR和W estern b lot结果表明重组质粒构建正确并可在mRNA和蛋白水平的表达,ELISA结果表明以重组质粒pVAX1-sig-LTB-lZP3-C3d3免疫诱导的特异性IgG、IgA的水平明显高于对照组(P<0.01)。抗生育实验表明该疫苗免疫的小鼠平均生仔数与对照组相比差异极显著(P<0.01)。结论:构建的草原兔尾鼠卵透明带3DNA疫苗重组质粒pVAX1-sig-LTB-lZP3-C3d3可高效地激发小鼠特异性免疫应答,提高抗生育的效果。
基金supported by the National Natural Science Foundation of China (No.60968002)the Program for New Century Excellent Talentsin University of China (No. NCET-05-0897)the Scientific Research Project for Universities in Xinjiang (No. XJEDU2006I10)
文摘A novel porous silicon based optical Fabry-Perot cavity structure is prepared as a label-free immunosensor platform for detecting antigen-antibody.The lagurus zona pelluciad 3(LZP3) and the specificity of the polyclonal anti-LZP3 antibodies are employed in our laboratory as the target and the probe,respectively.Firstly,the antibodies are immobilized to the porous silicon optical cavity using silanization and glutaraldehyde(GA) chemistry.And then,after the antigen-antibody reaction,it is monitored that the red shift of the reflection spectrum of the immunosensor increases with the antigen concentration.This research also plays a potential role for the extensive applications in immunoassay.
