[1]雷明明,邵西兵,施振旦.鸽子瘦素基因成熟肽重组蛋白的表达及多克隆抗体的制备[J].江苏农业学报,2017,(01):155-158.[doi:10.3969/j.issn.1000-4440.2017.01.025 ]
　LEI Ming-ming,SHAO Xi-bing,SHI Zhen-dan.Cloning of pigeon leptin mature peptide cDNA sequence and preparations of the recombinant protein and polyclonal antibody[J].,2017,(01):155-158.[doi:10.3969/j.issn.1000-4440.2017.01.025 ]
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鸽子瘦素基因成熟肽重组蛋白的表达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:
2017年01期
页码:
155-158
栏目:
畜牧兽医·水产养殖
出版日期:
2017-02-28

文章信息/Info

Title:
Cloning of pigeon leptin mature peptide cDNA sequence and preparations of the recombinant protein and polyclonal antibody
作者:
雷明明1; 邵西兵2; 施振旦1
（1. 江苏省农业科学院畜牧研究所,动物品种改良与繁育重点实验室，江苏南京210014；2. 广东省家禽研究所，广东广州510643）
Author(s):
LEI Ming-ming1; SHAO Xi-bing2; SHI Zhen-dan1
(1. Laboratory of Animal Breed Improvement and Reproduction，Institute of Animal Science Jiangsu Academy of Agricultural Science，Nanjing 210014, China；2. Institute of Guangdong Province Poultry Technology, Guangzhou 510520, China)
关键词:
鸽子瘦素; 成熟肽序列; 克隆; 表达; 多克隆抗体
Keywords:
pigeon leptin; mature peptide; cloning; expression; polyclonal antibody
分类号:
S836.2
DOI:
10.3969/j.issn.1000-4440.2017.01.025
文献标志码:
A
摘要:
根据鸽子瘦素基因（leptin）编码区序列（GenBank：HG797022）设计并合成鸽子leptin基因编码胞外域成熟肽的cDNA序列，然后将这段序列克隆到表达载体pGEX-4T-1的EcoR I与Not I酶切位点之间，构建重组表达载体(pGEX-4T-1-leptin)并将该载体转化到大肠杆菌Arctic Express中。转化菌经IPTG诱导后表达了分子量为44 270的鸽子leptin重组蛋白。经镍柱亲和层析纯化后的重组蛋白质与矿物油佐剂混合，免疫日本大耳白兔，制备抗leptin多克隆抗体。经过4次免疫之后，获得高效价的抗leptin多克隆抗体。
Abstract:
According to the pigeon leptin gene mature peptide sequence (GenBank：HG797022), the leptin gene were synthesized and cloned. The synthesis leptin gene was cloned into the sites of the plasmid pGEX-4T-1 between EcoR I and Not I to generate the expression vector pGEX-4T-1-leptin, which was further transformed into bacteria Arctic Express. The transformed bacteria were induced to produce recombinant leptin proteins with the expected molecular mass of 44 270 by IPTG. The recombinant leptin were purified by 50% Ni-NTA agrose chromatography and homogenized into mineral oil adjuvant. The mixers were used to immunize Japanese big ear rabbit to produce anti-leptin antibodies. Antisera with high anti-leptin titers were obtained after four immunizations.

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相似文献/References:

[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.[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,(01):1105.[doi:doi:10.3969/j.issn.1000-4440.2015.05.025]

备注/Memo

备注/Memo:
收稿日期：2016-03-14 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，副研究员，主要从事动物遗传与繁殖内分泌的研究。(Tel) 02584390772; (E-mail)mm0529@163.com 通讯作者：施振旦，(E-mail)zdshi@mail.jaas.ac.cn
更新日期/Last Update: 2017-04-12