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重组抗原CP_4-EPSPS的表达及免疫学活性研究 被引量:4
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作者 唐霓 Yu Xiang +4 位作者 Helen Ke zhihui yao 敬凌霞 黄爱龙 郑建 《重庆医科大学学报》 CAS CSCD 2006年第1期10-13,共4页
目的:表达并纯化重组CP4-EPSP合成酶(CP4-EPSPS),研究重组CP4-EPSPS的免疫学活性;为进一步制备单克隆抗体和开发检测CP4-EPSPS的快速诊断试剂奠定基础。方法:诱导表达含转基因大豆CP4-EPSPS的重组载体pET-EPSPS,经Ni-NTA琼脂糖树脂亲和... 目的:表达并纯化重组CP4-EPSP合成酶(CP4-EPSPS),研究重组CP4-EPSPS的免疫学活性;为进一步制备单克隆抗体和开发检测CP4-EPSPS的快速诊断试剂奠定基础。方法:诱导表达含转基因大豆CP4-EPSPS的重组载体pET-EPSPS,经Ni-NTA琼脂糖树脂亲和层析纯化,以ELISA、胶体金试条、W estern杂交鉴定其免疫活性。结果:重组CP4-EPSPS以包涵体表达为主,上清表达量极低,但胶体金试纸条检测发现上清中CP4-EPSPS抗原活性强;复性包涵体的抗原性与上清的抗原性明显不同。采用扩大培养量、缩小超声破碎重悬体积的方法,从上清中纯化CP4-EPSPS获得成功,ELISA检测此重组抗原免疫小鼠血清抗体滴度可达到1:625,000;W estern杂交证实重组抗原免疫血清与CP4-EPSPS标准品有较好的免疫反应性。结论:包涵体的简单复性方法难以恢复其关键的抗原决定簇的免疫原性;运用胶体金试纸条检测重组CP4-EPSPS活性,灵敏度高,简单快捷,对CP4-EPSPS特异检测相关的关键性抗原决定簇的甄别有重要作用,可指导重组抗原的纯化和在免疫学研究、特别是在单克隆抗体开发中的应用。 展开更多
关键词 CP4-EPSPS 快速检测 亲和层析 纯化
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A biological membrane-based novel excisional wound-splinting model in mice (With video)
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作者 zhihui yao Yong Huang +2 位作者 Gaoxing Luo Jun Wu Weifeng He 《Burns & Trauma》 SCIE 2014年第4期196-200,共5页
Rodents have robust wound healing mechanism compared to other animal species.The major mechanisms of wound healing diff er between rodents and humans.In humans,wound healing primarily depends on re-epithelialization a... Rodents have robust wound healing mechanism compared to other animal species.The major mechanisms of wound healing diff er between rodents and humans.In humans,wound healing primarily depends on re-epithelialization and granulation tissue(GT)formation,whereas wound contraction is more important during rodent wound closure.In this study,we described a novel excisional wound-splinting model in mice with a new biological membrane to imitate wound healing in humans.In this model,wound contraction can be eff ectually prevented,and the extent of re-epithelialization and the amount of granulation tissue can be determined easily.Furthermore,the harvested tissues can be analyzed with diff erent methods according to the research aim.In conclusion,we have developed a biological membrane-based,novel,excisional wound-splinting model in mice that has unique advantages for wound healing research compared with the conventional animal model. 展开更多
关键词 Animal model biological membrane RE-EPITHELIALIZATION wound healing
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