| 重阳木花粉相关变应原profilin基因的克隆、表达及免疫学鉴定 |
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| 引用本文： | 姚敏,刘志刚,邬玉兰,孟光,李春林.重阳木花粉相关变应原profilin基因的克隆、表达及免疫学鉴定[J].现代免疫学,2008,28(5). |
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| 作者姓名： | 姚敏 刘志刚 邬玉兰 孟光 李春林 |
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| 作者单位： | 姚敏,孟光,李春林,YAO Min,MENG Guang,LI Chun-lin(海口市人民医院检验科、耳鼻喉科,海口,570208);刘志刚,邬玉兰,LIU Zhi-gang,WU Yu-lan(深圳大学过敏反应与免疫学研究所,深圳,518060) |
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| 基金项目： | 国家自然科学基金，广东省科技重点专项，广东省深圳市科技计划，海南省自然科学基金计划项目 |
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| 摘 要： | 克隆、表达和纯化重阳木花粉相关变应原profilin基因,并对其免疫学活性进行鉴定。采用RT-PCR和3-’RACE技术获得整个profilin基因的开放阅读框,将其与PET28a载体连接并转化大肠杆菌E.cdiBL21(DE3)进行诱导表达,通过Ni2+亲和层析柱对重组蛋白进行纯化,采用Western blot检测其IgE结合活性。克隆获得了profilin的全长基因,开放阅读框为396个碱基(包括终止密码子),编码131个氨基酸。成功地构建了原核表达载体,并在大肠杆菌中大量地表达了profi-lin,纯化后的重组蛋白进行免疫印迹,结果显示重阳木花粉过敏患者血清对重组profilin反应呈阳性。
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| 关 键 词： | 重阳木 花粉过敏原 肌动蛋白抑制蛋白 重组过敏原 |
Cloning and expression of gene encoding pollen allergen profilin protein from Bischofia polycarpa and its immuological activities |
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| YAO Min,LIU Zhi-gang,WU Yu-lan,MENG Guang,LI Chun-lin.Cloning and expression of gene encoding pollen allergen profilin protein from Bischofia polycarpa and its immuological activities[J].Current Immunology,2008,28(5). |
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| Authors: | YAO Min LIU Zhi-gang WU Yu-lan MENG Guang LI Chun-lin |
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| Abstract: | The complete sequence of the open reading frame(ORF) was obtained by RT-PCR and 3'RACE technique,linked to expression vector pET28a and then transformed to E.coli BL21(DE3) for induction expression.The recombinant protein expressed was identified through Ni+ affinity chromatography and its IgE-binding activity was assayed by Western blotting.It was found that the complete sequence of ORF for profiling gene consisted of 396 base pairs(including the stop codon) coding 131 amino acids.In addition,the recombinant profilin protein could react specifically with sera of patients allergic to profilin protein from B.polycarpa indicating the specific IgE-binding activity of the recombinant profiling protein. |
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| Keywords: | Bischofia polycarpa pollen pollen allergy profilin recombinant allergy |
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