| 斑马鱼ANF基因的克隆及多克隆抗体的制备 |
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| 引用本文： | 李竞超,吴坚,梁艳,莫小阳,吴秀山. 斑马鱼ANF基因的克隆及多克隆抗体的制备[J]. 激光生物学报, 2011, 20(5): 645-649. DOI: 10.3969/j.issn.1007-7146.2011.05.014 |
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| 作者姓名： | 李竞超 吴坚 梁艳 莫小阳 吴秀山 |
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| 作者单位： | 湖南师范大学蛋白质化学及鱼类发育生物学教育部重点实验室,心脏发育研究中心,湖南长沙410081 |
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| 基金项目： | 国家自然科学基金项目(No.31071999,30771 170,30971 105,30971663,30871340,30671053,30970425,30900851); 湖南省自然科学基金项目(10JJ1006) |
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| 摘 要： | 心房钠尿肽ANF是钠尿肽家族中的一员,是心肌细胞分泌的一种循环激素,主要在心房组织分泌。生物信息学分析表明,斑马鱼ANF基因开放阅读框全长321bp,编码106个氨基酸,PCR扩增获得斑马鱼ANF基因全长。将所得的PCR片段插入原核表达载体pGEX-4T-1中,并将重组质粒(pGEX-4T-1-ANF)转化大肠杆菌BL21。通过IPTG诱导表达GST-ANF融合蛋白,经GST亲和层析法纯化后,免疫新西兰大白兔制备了多克隆抗体,Western blotting分析表明,制备的抗体具有良好的高效价性和特异性。
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| 关 键 词： | ANF基因 融合蛋白 多克隆抗体 |
Cloning and Polyclonal Antibody Preparation of ANF Gene in Zebrafish |
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| LI Jingchao,WU Jian,LIANG Yan,MO Xiaoyang,WU Xiushan. Cloning and Polyclonal Antibody Preparation of ANF Gene in Zebrafish[J]. ACTA Laser Biology Sinica, 2011, 20(5): 645-649. DOI: 10.3969/j.issn.1007-7146.2011.05.014 |
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| Authors: | LI Jingchao WU Jian LIANG Yan MO Xiaoyang WU Xiushan |
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| Affiliation: | LI Jingchao,WU Jian,LIANG Yan,MO Xiaoyang~*,WU Xiushan~* (The Center for Heart Development,Key Lab of MOE for Development Biology and Protein Chemistry,Hunan Normal University,Changsha 410081,Hunan,China) |
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| Abstract: | Atrial natriuretic peptide ANF is one member of natriuretic peptide family,which is a circulation excitatory autacoid secreted by myocardial cell,mainly secrete in cardiac atrium.Zebrafish ANF gene open reading frame has 321 bp,encodes 106 amino acids.The full-length of ANF gene in zebrafish was cloned by bioinformatics combined with PCR technique.We inserted the PCR fragment into prokaryotic expression vector pGEX-4T-1,and then transformed the recombinant plasmid(pGEX-4T-1-ANF) into BL21.The GST-ANF fusion... |
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| Keywords: | ANF fusion protein polytolonal antibody |
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