| 鸡蛋主要过敏原Gal d1片段基因的克隆、表达、纯化及免疫学鉴定 |
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| 引用本文： | 马艳梅,黄钟,刘志刚,闫浩,王和平,罗新萍,孔小丽. 鸡蛋主要过敏原Gal d1片段基因的克隆、表达、纯化及免疫学鉴定[J]. 免疫学杂志, 2010, 0(7) |
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| 作者姓名： | 马艳梅 黄钟 刘志刚 闫浩 王和平 罗新萍 孔小丽 |
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| 作者单位： | 深圳大学医学院;深圳大学医学院呼吸疾病国家重点实验室深圳大学变态反应分室;深圳市儿童医院内五科; |
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| 基金项目： | 深圳市科技计划项目(JC200903130238A) |
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| 摘 要： | 目的克隆、表达和纯化鸡蛋主要过敏原Gald1区基因,并对其免疫学活性进行鉴定。方法用生物信息学方法对鸡蛋主要过敏原Gald1基因进行抗原表位预测,设计特异性引物,用PCR的方法分别扩增Gald1N端和C端基因,将其分别连接到原核表达载体PET32a上。重组质粒转入大肠杆菌,用IPTG诱导表达,通过镍亲和柱层析法纯化重组蛋白,最后用Western-blotting检测重组蛋白的免疫原性。结果表达的蛋白均以可溶性为主,N端融合蛋白相对分子质量为28600,C端融合蛋白相对分子质量为26800,纯化的N端和C端蛋白均有较好的免疫原性,C端的免疫原性强于N端。结论成功地克隆和表达了鸡蛋主要变应原基因Gald1的N端和C端,为鸡蛋过敏的诊断和免疫治疗奠定了基础。
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| 关 键 词： | 鸡蛋过敏原 Gald1 克隆 表达 |
Cloning,expression and identification the DNA fragments of main allergen Gal d1 in Egg |
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| MA Yanmei,HUANG Zhong,LIU Zhigang,YAN Hao,WANG Heping,LIU Xinping,KONG Xiaoli Medical College,Shenzhen University,Shenzhen ,China. Cloning,expression and identification the DNA fragments of main allergen Gal d1 in Egg[J]. Immunological Journal, 2010, 0(7) |
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| Authors: | MA Yanmei HUANG Zhong LIU Zhigang YAN Hao WANG Heping LIU Xinping KONG Xiaoli Medical College Shenzhen University Shenzhen China |
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| Affiliation: | MA Yanmei,HUANG Zhong,LIU Zhigang,YAN Hao,WANG Heping,LIU Xinping,KONG Xiaoli Medical College,Shenzhen University,Shenzhen 518060,China |
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| Abstract: | To clone,express and identify the fragments of the main allergen Gal d1 in egg,the epitopes of Gal d1 gene were compared by bioinformatic methods,and the conservative domains among the sequences were used for designing PCR primers.The N terminal gene and C terminal gene of Gal d1 were amplified by PCR,and then conjuncted them to the vectors for cloning.And positive clones were ligated into pET32a for expression.After induced by IPTG,recombinant proteins were purified by metal(Ni2+) chelating affinity chroma... |
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| Keywords: | Egg allergen Gal d1 Clone Expression |
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