| 蛔虫过敏原ABA-1融合基因在大肠杆菌中的表达及鉴定 |
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| 引用本文： | 于超生,文忠,邹泽红,陈惠芳,陶爱林. 蛔虫过敏原ABA-1融合基因在大肠杆菌中的表达及鉴定[J]. 中国组织工程研究与临床康复, 2011, 15(20): 3679-3682. DOI: 10.3969/j.issn.1673-8225.2011.20.017 |
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| 作者姓名： | 于超生 文忠 邹泽红 陈惠芳 陶爱林 |
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| 作者单位： | 1. 南方医科大学珠江医院耳鼻咽喉-头颈外科,广东省广州市,510282
2. 广州医学院第二附属医院过敏反应与临床免疫重点实验室,呼吸疾病国家重点实验室,广东省广州市510260 |
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| 基金项目： | 课题受国家科技重大专项重大课题，重点课题，国家自然科学基金项，广州市教育系统科研创新学术团队 |
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| 摘 要： | 背景:利用蛔虫基因融合技术,可将过敏原ABA-1的主要基因进行融合。目的:利用大肠杆菌表达系统重组表达蛔虫主要过敏原ABA-1融合蛋白。方法:从Gene Bank和Protein Database中获取蛔虫主要过敏原ABA-1基因和蛋白序列,选定其中的ABA-1B1,ABA-1A2与ABA-1A1基因重组为融合基因BAA,经密码子优化后,全基因合成目的基因BAA并构建于表达载体PET-44a上,经KCM法转化入大肠杆菌JM109中进行克隆,PET-44a/BAA经NdeⅠ和PstⅠ双酶切及DNA测序正确后,转化入大肠杆菌Rosetta Blue TM中,经IPTG诱导表达。表达蛋白经Ni柱亲和层析纯化后,通过Western blot和氨基酸测序鉴定目的蛋白。结果与结论:大肠杆菌的融合蛋白BAA表达量约占总蛋白的40%,纯化后蛋其白纯度可达90%左右。Western Blot结果显示在相对分子质量45000处可见一特异目的条带,氨基酸测序显示N端15个氨基酸与目的蛋白完全相同。结果证实,融合蛋白BAA在大肠杆菌中得到高效的表达。
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| 关 键 词： | 蛔虫 过敏原 ABA-1 密码子优化 组织构建 |
Expression and identification of chimeric protein truncated from Ascaris allergen ABA-1 |
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| Yu Chao-sheng,Wen Zhong,Zou Ze-hong,Chen Hui-fang,Tao Ai-lin. Expression and identification of chimeric protein truncated from Ascaris allergen ABA-1[J]. Journal of Clinical Rehabilitative Tissue Engineering Research, 2011, 15(20): 3679-3682. DOI: 10.3969/j.issn.1673-8225.2011.20.017 |
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| Authors: | Yu Chao-sheng Wen Zhong Zou Ze-hong Chen Hui-fang Tao Ai-lin |
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| Affiliation: | Yu Chao-sheng1,Wen Zhong1,Zou Ze-hong2,Chen Hui-fang2,Tao Ai-lin2 1Department of Otolaryngology-Head and Neck Surgery,Zhujiang Hospital,Southern Medical University,Guangzhou 510282,Guangdong Province,China,2Academic Key Laboratory of Allergy and Clinical Immunity,State Key Laboratory of Respiratory Diseases,the Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260 |
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| Abstract: | BACKGROUND:Serious allergic reaction could be caused by Ascaris anaphylactogen ABA-1 protein (Ascaris Body fluid allergen 1).OBJECTIVE:To express the chimeric protein truncated from Ascais major allergen ABA-1 in Escherichia coli.METHODS:The chimeric gene,obtained from GeneBank and Protein Database and named as BAA hereafter,was constructed by tailoring the coding fragments of Ascaris main allergen ABA-1,i.e.,ABA-1B1,ABA-1A2 and ABA-1A1.The fusion gene BAA was synthesized and cloned into expression vector P... |
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| Keywords: | ABA-1 |
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