构建人基因组的高分辨遗传图和更精细的互补物理图,收集按次序排列的 DNA 克隆,测定一个参考基因组核苷酸全序列,确定所有基因的位置,以此应用于生物和医学,平行地研究模式生物以充分而全面地了解人类基因组,这就是人类基因组研究计划...构建人基因组的高分辨遗传图和更精细的互补物理图,收集按次序排列的 DNA 克隆,测定一个参考基因组核苷酸全序列,确定所有基因的位置,以此应用于生物和医学,平行地研究模式生物以充分而全面地了解人类基因组,这就是人类基因组研究计划的目标。展开更多
目的了解腺病毒载体转染人HO-1基因对体外培养的成人胰岛的作用,探索基因治疗在胰岛移植中的潜在应用价值。方法将成人胰岛分离纯化后分为3组:转染人HO-1基因组(Ad-HO-1组)、转染EGFP基因组(Ad-EGFP组)及对照组,采用携带人HO-1基因及EGF...目的了解腺病毒载体转染人HO-1基因对体外培养的成人胰岛的作用,探索基因治疗在胰岛移植中的潜在应用价值。方法将成人胰岛分离纯化后分为3组:转染人HO-1基因组(Ad-HO-1组)、转染EGFP基因组(Ad-EGFP组)及对照组,采用携带人HO-1基因及EGFP基因的腺病毒作为载体对体外培养的成人胰岛进行转染,通过形态学观察、胰岛素释放试验及肿瘤坏死因子(TNF)α及放线菌酮诱导48 h后流式细胞仪检测凋亡。结果Ad-HO-1组的胰岛在高糖刺激下胰岛素分泌量为270 m IU/L±89 m IU/L,高于对照组(182 m IU/L±59 m IU/L)和Ad-EGFP组(189 m IU/L±88 m IU/L,P<0.05);诱导后Ad-HO-1组凋亡细胞的发生率为63%±11%,对照组为91%±11%,两组比较,P<0.01。结论使用腺病毒作为载体对体外培养的成人胰岛转染HO-1基因能够增加胰岛的抗凋亡能力、促进胰岛素的分泌。展开更多
目的评价限制性片段长度多态性（Restriction fragment length polymorphism，RFLP）方法检测α和a交配型位点内GEF1α/a基因片段在鉴定新生和格特隐球菌种、变种、基因型和交配型中的作用。方法筛选交配型位点内的GEF1α/a基因进行PCR...目的评价限制性片段长度多态性（Restriction fragment length polymorphism，RFLP）方法检测α和a交配型位点内GEF1α/a基因片段在鉴定新生和格特隐球菌种、变种、基因型和交配型中的作用。方法筛选交配型位点内的GEF1α/a基因进行PCR—RFLP分析。根据各基因型和交配型参考株的GEF1α/a基因保守序列设计引物，扩增受试新生和格特隐球菌GEF1α/a基因部分片段。利用DNAMAN和Vector NTI软件进行序列比对、限制性图谱分析、限制性内切酶的筛选和模拟电泳。选择EcoT14 Ⅰ和Hap Ⅱ限制性内切酶分别对125株新生和格特隐球菌的GEF1α/a基因扩增片段进行RFLP分型。结果所有受试的82株新生隐球菌和43株格特隐球菌均扩增出1300bp大小片段，而罗伦隐球菌、白念珠菌、热带念珠菌、近平滑念珠菌、克柔念珠菌、光滑念珠菌、阿萨希毛孢子菌、烟曲霉和黄曲霉参考株均扩增阴性。RFLP分型准确鉴定所有125株新生和格特隐球菌的种、变种、基因型和交配型。结论针对GEF1α/a基因片段的PCR—RFLP方法特异性高、稳定性好，适用于新生和格特隐球菌种、变种、基因型和交配型的同步和快速鉴定以及进一步的分子流行病学分析。展开更多
构建了含抗人端粒酶逆转录酶单链抗体基因ScFv-hTERT的植物表达载体p1304-SH,通过农杆菌介导的叶盘法转化烟草.转基因烟草植株叶片总DNA的PCR,Southern b lot检测结果表明,ScFv-hTERT基因已整合进了转基因烟草植株基因组中;RT-PCR,SDS-P...构建了含抗人端粒酶逆转录酶单链抗体基因ScFv-hTERT的植物表达载体p1304-SH,通过农杆菌介导的叶盘法转化烟草.转基因烟草植株叶片总DNA的PCR,Southern b lot检测结果表明,ScFv-hTERT基因已整合进了转基因烟草植株基因组中;RT-PCR,SDS-PAGE分析证实目的基因已在烟草叶片中成功表达,竞争性ELISA的结果表明,表达的重组抗体与其抗原有良好的结合活性.展开更多
The effect of antibacterial peptide CM4 of Bombyx mori against E. coll K12 was investigated using scanning electron microscopy(SEM) and transmission electron microscopy (TEM). The ultrastructural changes of E. coli K1...The effect of antibacterial peptide CM4 of Bombyx mori against E. coll K12 was investigated using scanning electron microscopy(SEM) and transmission electron microscopy (TEM). The ultrastructural changes of E. coli K12 were observed by the challenge of the purified antibacterial peptide CM4. The results showed that the antibacterial peptide caused a series of pathological changes on E. coli. SEM and TEM revealed aggregates of bacteria and SEM revealed wrin-kled bacterial surfaces in the early stage. Thereafter, plasmolysis was observed with irregular holes appearing in the two ends of bacteria and the cytoplasmic contents of the cells leaking out. Finally, bacteria became empty vesicles and disintegrated into small fragments subsequently. Comparatively, the bacterial membrane was normal and the bacterial structure remained intact in the control group.展开更多
文摘目的了解腺病毒载体转染人HO-1基因对体外培养的成人胰岛的作用,探索基因治疗在胰岛移植中的潜在应用价值。方法将成人胰岛分离纯化后分为3组:转染人HO-1基因组(Ad-HO-1组)、转染EGFP基因组(Ad-EGFP组)及对照组,采用携带人HO-1基因及EGFP基因的腺病毒作为载体对体外培养的成人胰岛进行转染,通过形态学观察、胰岛素释放试验及肿瘤坏死因子(TNF)α及放线菌酮诱导48 h后流式细胞仪检测凋亡。结果Ad-HO-1组的胰岛在高糖刺激下胰岛素分泌量为270 m IU/L±89 m IU/L,高于对照组(182 m IU/L±59 m IU/L)和Ad-EGFP组(189 m IU/L±88 m IU/L,P<0.05);诱导后Ad-HO-1组凋亡细胞的发生率为63%±11%,对照组为91%±11%,两组比较,P<0.01。结论使用腺病毒作为载体对体外培养的成人胰岛转染HO-1基因能够增加胰岛的抗凋亡能力、促进胰岛素的分泌。
文摘构建了含抗人端粒酶逆转录酶单链抗体基因ScFv-hTERT的植物表达载体p1304-SH,通过农杆菌介导的叶盘法转化烟草.转基因烟草植株叶片总DNA的PCR,Southern b lot检测结果表明,ScFv-hTERT基因已整合进了转基因烟草植株基因组中;RT-PCR,SDS-PAGE分析证实目的基因已在烟草叶片中成功表达,竞争性ELISA的结果表明,表达的重组抗体与其抗原有良好的结合活性.
文摘The effect of antibacterial peptide CM4 of Bombyx mori against E. coll K12 was investigated using scanning electron microscopy(SEM) and transmission electron microscopy (TEM). The ultrastructural changes of E. coli K12 were observed by the challenge of the purified antibacterial peptide CM4. The results showed that the antibacterial peptide caused a series of pathological changes on E. coli. SEM and TEM revealed aggregates of bacteria and SEM revealed wrin-kled bacterial surfaces in the early stage. Thereafter, plasmolysis was observed with irregular holes appearing in the two ends of bacteria and the cytoplasmic contents of the cells leaking out. Finally, bacteria became empty vesicles and disintegrated into small fragments subsequently. Comparatively, the bacterial membrane was normal and the bacterial structure remained intact in the control group.
