| 冬虫夏草指标蛋白IP4的cDNA全长克隆及抗原决定簇预测研究 |
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| 引用本文： | 张涵,童芯锌,王芳,王艺璇,彭成,国锦琳. 冬虫夏草指标蛋白IP4的cDNA全长克隆及抗原决定簇预测研究[J]. 中草药, 2018, 49(20): 4864-4869 |
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| 作者姓名： | 张涵 童芯锌 王芳 王艺璇 彭成 国锦琳 |
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| 作者单位： | 成都中医药大学中药材标准化教育部重点实验室中药资源系统研究与开发利用国家重点实验室培育基地 |
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| 基金项目： | 国家自然科学基金资助项目（30801522，81373920）；四川省中医药管理局川产道地药材大品种项目；成都市科技局科技惠民项目资助 |
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| 摘 要： | 目的克隆获取冬虫夏草指标蛋白氰酸酶(IP4)的c DNA全长及对其抗原表位预测。方法结合课题组前期采用蛋白组学法筛选得到的冬虫夏草指标性蛋白IP4和冬虫夏草转录组数据,挖掘IP4 m RNA序列,并克隆IP4 c DNA全长,通过NCBI结构分析服务系统对IP4进行保守区和功能区分析,利用DNAMAN 6.0和DNAStar软件分别进行序列比对和抗原表位分析。结果从转录组数据中挖掘到IP4的2个可变剪接体,属于内含子保留的可变剪接。RT-PCR结果显示,IP4 c DNA全长465 bp,编码154 aa(氨基酸)。IP4蛋白的保守区和主要催化活性位点位于87～154 aa,该区域为氰酸酶家族保守核心区域和二聚体结合区域,DNAMAN 6.0分析结果表明1～39 aa和55～81 aa区具有较高种属特异性,DNAStar预测IP4蛋白的抗原表位区主要分布于25～90 aa。结论最终确定IP4的25～39 aa和55～81 aa为最佳抗原特异区,为后续规模化制备IP4蛋白特异抗原区及免疫法鉴定冬虫夏草奠定基础。
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| 关 键 词： | 冬虫夏草 指标蛋白 cDNA克隆 DNAStar 抗原区分析 |
| 收稿时间： | 2018-04-30 |
cDNA cloning and prediction of antigenic determinants of protein marker IP4 in Ophiocordyceps sinensis |
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| ZHANG Han,TONG Xin-xin,WANG Fang,WANG Yi-xuan,PENG Cheng and GUO Jin-lin. cDNA cloning and prediction of antigenic determinants of protein marker IP4 in Ophiocordyceps sinensis[J]. Chinese Traditional and Herbal Drugs, 2018, 49(20): 4864-4869 |
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| Authors: | ZHANG Han TONG Xin-xin WANG Fang WANG Yi-xuan PENG Cheng GUO Jin-lin |
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| Affiliation: | State Key Laboratory Breeding Base of TCM Resources Research and Development, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine of Ministry of Education, Chengdu 611137, China,State Key Laboratory Breeding Base of TCM Resources Research and Development, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine of Ministry of Education, Chengdu 611137, China,State Key Laboratory Breeding Base of TCM Resources Research and Development, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine of Ministry of Education, Chengdu 611137, China,State Key Laboratory Breeding Base of TCM Resources Research and Development, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine of Ministry of Education, Chengdu 611137, China,State Key Laboratory Breeding Base of TCM Resources Research and Development, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine of Ministry of Education, Chengdu 611137, China and State Key Laboratory Breeding Base of TCM Resources Research and Development, Key Laboratory of Standardization of Chinese Herbal Medicine, Chengdu University of Traditional Chinese Medicine of Ministry of Education, Chengdu 611137, China |
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| Abstract: | |
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| Keywords: | Ophiocordyceps sinensis (Berk.) Sacc. protein marker cDNA clone DNAStar antigenic site analysis |
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