[1]雷明明,吴思谦,李孝伟,等.鸡瘦素受体胞外域成熟肽重组蛋白质的表达以及多克隆抗体的制备[J].江苏农业学报,2015,(05):1105-1109.[doi:doi:10.3969/j.issn.1000-4440.2015.05.025]
　LEI Ming-ming,WU Si-qian,LI Xiao-wei,et al.Expression of recombinant protein of chicken leptin receptor extracellular domain mature peptide and the preparation of its polyclonal antibody[J].,2015,(05):1105-1109.[doi:doi:10.3969/j.issn.1000-4440.2015.05.025]
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鸡瘦素受体胞外域成熟肽重组蛋白质的表达以及多克隆抗体的制备()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年05期
页码:
1105-1109
栏目:
畜牧兽医·水产养殖
出版日期:
2015-10-31

文章信息/Info

Title:
Expression of recombinant protein of chicken leptin receptor extracellular domain mature peptide and the preparation of its polyclonal antibody
作者:
雷明明1; 吴思谦2; 李孝伟2; 施振旦1
（1.江苏省农业科学院畜牧研究所动物品种改良和繁育重点实验室，江苏南京210014；2.华南农业大学动物科学学院，广东广州510642）
Author(s):
LEI Ming-ming1; WU Si-qian2; LI Xiao-wei2; SHI Zhen-dan1
(1.Laboratory of Animal Breed Improvement and Reproduction, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences，Nanjing 210014, China；2.College of Animal Science, South China Agricultural University, Guangzhou 510642, China)
关键词:
瘦素受体; 成熟肽序列; 克隆和表达; 多克隆抗体
Keywords:
leptin receptor; mature peptide; cloning and expression; polyclonal antibody
分类号:
S831.2
DOI:
doi:10.3969/j.issn.1000-4440.2015.05.025
文献标志码:
A
摘要:
根据鸡瘦素受体基因（LEPR）编码区序列（GenBank：NM_AB033383）设计并合成2对引物，以鸡肝脏cDNA为模板扩增鸡LEPR胞外域2段cDNA序列。然后将这2段序列克隆到表达载体pRSET-A的BamH I和Hind III酶切位点之间，构建重组表达载体(pR-LEPR1和pR-LEPR2 )并转化大肠杆菌E.coli BL21(DE3)。转化菌在IPTG诱导后分别表达了分子量为 2.62×104的 LEPR1和分子量为 2.86×104的LEPR2重组蛋白质。经凝胶Ni-NTA纯化后的重组蛋白质与矿物油佐剂混合免疫生长鸡，经过3次免疫之后，获得高效价的抗LEPR1和抗LEPR2抗血清。
Abstract:
Two pairs of primers designed based on the chicken leptin receptor gene (LEPR) mature peptide sequence (GenBank：NM_AB033383) were used to amplify two cDNA sequence fragments of chicken LEPR extracellular domains (ECD) mature peptides with chicken liver cDNA as the template. The two cDNA sequence fragments were cloned into the BamH I and Hind III sites of the plasmid pRSET-A to generate the expression vector pR-LEPR1 and pR-LEPR2, which were further transformed into bacterium Escherichia coli BL21(DE3). The transformed bacteria were induced to produce recombinant proteins LEPR1 and LEPR2 with the expected molecular mass of 2.62×104 and 2.86×104 by IPTG. The recombinant LEPR1 and LEPR2 were purified and mixed into mineral oil adjuvant to immunize growing chickens to produce anti-LEPR1 and anti-LEPR2 antibodies. Antisera with high anti-LEPR1 and anti-LEPR2 titers were obtained after three immunizations.〖JP〗

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备注/Memo

备注/Memo:
收稿日期：2015-03-05 基金项目：国家自然科学基金项目(31501946);江苏省自然科学基金项目（BK20150544） 作者简介：雷明明（1977-），女，湖北赤壁人，博士，研究方向为动物遗传与繁殖内分泌。( Tel ) 025-84390772; ( E-mail )mm0529@163.com 通讯作者：施振旦（1964-），(Tel)025-84390956;(E-mail)zdshi@mail.jaas.ac.cn
更新日期/Last Update: 2015-10-31