目的探讨HBV相关性肾炎(hepatitis B virus associated glomerulonephritis,HBV-GN)易感性与维生素D受体(vitamin D receptor,VDR)基因BsmⅠ及FokⅠ酶切位点多态性的关系。方法应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析技...目的探讨HBV相关性肾炎(hepatitis B virus associated glomerulonephritis,HBV-GN)易感性与维生素D受体(vitamin D receptor,VDR)基因BsmⅠ及FokⅠ酶切位点多态性的关系。方法应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析技术检测VDR基因BsmⅠ及FokⅠ的多态性在40例HBV-GN患者、34例慢性乙肝患者及46例健康对照者中的分布,并进行基因型分析。结果 HBV-GN组FF基因型分布明显低于正常对照组及慢性乙肝组(χ2=9.084、10.254,P=0.0025、0.0014),HBV-GN组F/f等位基因频率分布和对照组及慢性乙肝组比较无统计学意义(P>0.05);慢性乙肝组FokⅠ位点基因型和等位基因频率分布与正常对照组比较差异不显著(P>0.05)。HBV-GN组和乙型肝炎组Bb基因型分布明显高于正常对照组(χ2=8.9451、0.435,P=0.0027、0.0012),两组B等位基因分布明显高于对照组(χ2=7.9001、0.329,P=0.00490、.0013);HBV-GN和慢性乙肝组间比较差异不显著。结论 VDR基因多态性可能与乙肝相关性肾炎及HBV感染相关联。展开更多
AIM To construct the recombinant of HDVcDNA and HBV-specific ribozyme gene byrecombinant PCR in order to use HDV as atransporting vector carrying HBV-specificribozyme into liver cells for inhibiting thereplication of ...AIM To construct the recombinant of HDVcDNA and HBV-specific ribozyme gene byrecombinant PCR in order to use HDV as atransporting vector carrying HBV-specificribozyme into liver cells for inhibiting thereplication of HBV.METHODS We separately cloned the ribozyme（RZ）gene and recombinant DVRZ（comprisingHDV cDNA and HBV-specific ribozyme gene）intothe downstream of T7 promoter of pTAdv-Tvector and studied the in vitro cleavage activityof their transcripts（rRZ,rDVRZ）on target RNA（rBVCF）from in vitro transcription of HBV Cgene fragment（BVCF）.RESULTS Both the simple（rRZ）and therecombinant ribozyme rDVRZ could efficientlycatalyze the cleavage of target RNA（rBVCF）under different temperatures（37℃,42℃ and55℃）and Mg<sup>2+</sup>concentrations（10 mmol/L,15 mmol/L and 20 mmol/L）and their catalyticactivity tended to increase as the temperaturewas rising.But the activity of rRZ was evidentlyhigher than that of rDVRZ.CONCLUSION The recombinant of HDV cDNAand ribozyme gene had the potential of beingfurther explored and used in gene therapy of HBVinfection.展开更多
Amplified HDV fragment by RT-PCR was cloned into T vector and PQE_{31} plasmid, which established a prokaryotic expression vector in E coli M_{15} The recombinant protein was purified by Ni-NTA column affinity chromat...Amplified HDV fragment by RT-PCR was cloned into T vector and PQE_{31} plasmid, which established a prokaryotic expression vector in E coli M_{15} The recombinant protein was purified by Ni-NTA column affinity chromatography, and identified by Western blot and ELISA: HDV antigen was expressed efficiently and had a well antigenicity character. It can be used in HDV clinical diagnosis and epidemiology survey.展开更多
文摘目的探讨HBV相关性肾炎(hepatitis B virus associated glomerulonephritis,HBV-GN)易感性与维生素D受体(vitamin D receptor,VDR)基因BsmⅠ及FokⅠ酶切位点多态性的关系。方法应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析技术检测VDR基因BsmⅠ及FokⅠ的多态性在40例HBV-GN患者、34例慢性乙肝患者及46例健康对照者中的分布,并进行基因型分析。结果 HBV-GN组FF基因型分布明显低于正常对照组及慢性乙肝组(χ2=9.084、10.254,P=0.0025、0.0014),HBV-GN组F/f等位基因频率分布和对照组及慢性乙肝组比较无统计学意义(P>0.05);慢性乙肝组FokⅠ位点基因型和等位基因频率分布与正常对照组比较差异不显著(P>0.05)。HBV-GN组和乙型肝炎组Bb基因型分布明显高于正常对照组(χ2=8.9451、0.435,P=0.0027、0.0012),两组B等位基因分布明显高于对照组(χ2=7.9001、0.329,P=0.00490、.0013);HBV-GN和慢性乙肝组间比较差异不显著。结论 VDR基因多态性可能与乙肝相关性肾炎及HBV感染相关联。
基金Natural Science Foundation of Guangdong Province,No.940311.
文摘AIM To construct the recombinant of HDVcDNA and HBV-specific ribozyme gene byrecombinant PCR in order to use HDV as atransporting vector carrying HBV-specificribozyme into liver cells for inhibiting thereplication of HBV.METHODS We separately cloned the ribozyme（RZ）gene and recombinant DVRZ（comprisingHDV cDNA and HBV-specific ribozyme gene）intothe downstream of T7 promoter of pTAdv-Tvector and studied the in vitro cleavage activityof their transcripts（rRZ,rDVRZ）on target RNA（rBVCF）from in vitro transcription of HBV Cgene fragment（BVCF）.RESULTS Both the simple（rRZ）and therecombinant ribozyme rDVRZ could efficientlycatalyze the cleavage of target RNA（rBVCF）under different temperatures（37℃,42℃ and55℃）and Mg<sup>2+</sup>concentrations（10 mmol/L,15 mmol/L and 20 mmol/L）and their catalyticactivity tended to increase as the temperaturewas rising.But the activity of rRZ was evidentlyhigher than that of rDVRZ.CONCLUSION The recombinant of HDV cDNAand ribozyme gene had the potential of beingfurther explored and used in gene therapy of HBVinfection.
文摘Amplified HDV fragment by RT-PCR was cloned into T vector and PQE_{31} plasmid, which established a prokaryotic expression vector in E coli M_{15} The recombinant protein was purified by Ni-NTA column affinity chromatography, and identified by Western blot and ELISA: HDV antigen was expressed efficiently and had a well antigenicity character. It can be used in HDV clinical diagnosis and epidemiology survey.
