| 孔石莼凝集素蛋白基因的克隆与表达 |
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| 引用本文： | 郑璐平,谢荔岩,姚锦爱,钟伏弟,林奇英,吴祖建,谢联辉.孔石莼凝集素蛋白基因的克隆与表达[J].激光生物学报,2008,17(6). |
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| 作者姓名： | 郑璐平 谢荔岩 姚锦爱 钟伏弟 林奇英 吴祖建 谢联辉 |
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| 作者单位： | 1. 福建农林大学病毒研究所,福建,福州,350002;福建农林大学生物农药与化学生物学教育部重点实验室,福建,福州,350002
2. 福建农林大学病毒研究所,福建,福州,350002 |
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| 基金项目： | 福建省科技厅科技计划
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| 摘 要： | 根据孔石莼(Ulva pertusa)凝集素(Lectin)蛋白cDNA全长序列(GenBank登录号:AY433960)设计引物,以其总DNA为模板,采用PCR技术扩增蛋白DNA序列,经克隆、测序获得基因序列。结果表明,孔石莼凝集素蛋白(UPL)基因序列长约为670 bp,含有一个大小为56 bp的内含子。此外,设计带酶切位点的引物,以UPL-cDNA为模板,扩增其开放阅读框,并与表达载体pGEX-2T连接,构建原核表达载体pG2T-UPL,并在大肠杆菌BL21(DE3)中成功表达大小约为47 kD的目的蛋白。
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| 关 键 词： | 孔石莼 凝集素 基因克隆 原核表达 |
Cloning and Expression of Lectin Gene of Ulva pertusa |
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| ZHENG Lu-ping,XIE Li-yan,YAO Jin-ai,ZHONG Fu-di,LIN Qi-ying,WU Zu-jian,XIE Lian-hui.Cloning and Expression of Lectin Gene of Ulva pertusa[J].ACTA Laser Biology Sinica,2008,17(6). |
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| Authors: | ZHENG Lu-ping XIE Li-yan YAO Jin-ai ZHONG Fu-di LIN Qi-ying WU Zu-jian XIE Lian-hui |
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| Abstract: | Based on the known cDNA sequence of lectin protein gene of U.pertusa(GenBank accession number: AY433960),a pair of primers were designed and used to clone,and sequence the lectin protein gene with PCR technique by using the genomic DNA of U.pertusa as the template.The results showed that the lectin protein gene consisted of 670 bp,with a 56 bp intron.A pair of primers with restriction enzyme sites were designed to amplify the ORF using the genomic cDNA of U.pertusa as the template.The ORF was inserted into the expression vector pGEX-2T and a protein with molecular weight of 47 kD was successfully expressed in Escherichia coli BL21(DE3). |
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| Keywords: | cDNA |
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