盐穗木HcDMC1的原核表达和抗血清的制备

doi:10.7525/j.issn.1673-5102.2018.03.012

植物研究 ›› 2018, Vol. 38 ›› Issue (3): 399-405.doi: 10.7525/j.issn.1673-5102.2018.03.012

盐穗木HcDMC1的原核表达和抗血清的制备

张冀^1,2, 张丽丽^1,2, 张贝贝^1,2, 张富春^1,2

1. 新疆大学生命科学与技术学院, 乌鲁木齐 830046;
2. 新疆生物资源基因工程重点实验室, 乌鲁木齐 830046

收稿日期:2017-08-30 出版日期:2018-05-15 发布日期:2018-05-17
通讯作者: 张富春,E-mail:zfcxju@xju.edu.cn E-mail:zfcxju@xju.edu.cn
作者简介:张冀(1991-),男,硕士研究生,主要从事植物分子生物学研究。
基金资助:
新疆重点实验室专项资金资助项目（2014KL001）

Prokaryotic Expression of Halostachys caspica HcDMC1 and Preparation of Its Antiserum

ZHANG Ji^1,2, ZHANG Li-Li^1,2, ZHANG Bei-Bei^1,2, ZHANG Fu-Chun^1,2

1. College of Life Science and Technology, Xinjiang University, Urumqi 830046;
2. Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi 830046

Received:2017-08-30 Online:2018-05-15 Published:2018-05-17
Supported by:
Supported by Special Funds of Xinjiang Key Laboratory(2014KL001)

摘要/Abstract

摘要： DMC1是减数分裂过程中同源染色体配对和重组修复所必需的减数分裂特异蛋白。根据盐胁迫下盐穗木转录组数据库，克隆获得的盐穗木DNA损伤修复基因命名为HcDMC1。为深入分析盐穗木HcDMC1基因的耐盐功能，通过原核表达获得盐穗木HcDMC1的融合蛋白，用于免疫小鼠制备特异性的HcDMC1抗血清。结果表明，利用pET30a-HcDMC1能够在大肠杆菌BL21（DE3）中诱导表达融合蛋白His-HcDMC1，纯化融合蛋白His-HcDMC1的含量为1.0 mg·mL^-1。每次每只小鼠免疫接种50 μg融合蛋白，三次免疫接种后进行抗体效价及特异性检测。ELISA检测抗血清滴度约为1：400 000，Western Blot检测证明了抗血清的特异性。本研究制备的小鼠抗血清能够为盐穗木HcDMC1蛋白的功能鉴定和免疫检测提供实验材料。

关键词: 盐穗木, HcDMC1, 原核表达, 融合蛋白, 抗血清

Abstract: DMC1 is a necessary meiotic specific protein for homologous chromosome pairing and recombinant repair during meiosis. According to the transcriptome database of Halostachys caspica under the salt stress, the DNA damage repair gene DMC1 of H. caspica was cloned and named HcDMC1. In order to further investigate the salt tolerant function of HcDMC1, the HcDMC1 fusion protein obtained from prokaryotic expression was used to immunize mice to prepare the specific antiserum against HcDMC1. The expression of HcDMC1 protein could be induced in Escherichia coli by pET30a-HcDMC1 and the fusion protein His-HcDMC1 was purified. The content of His-HcDMC1 was 1 mg·mL^-1. Each mouse was immunized with 50 μg fusion protein for three time, the antibody titer and specificity were detected by ELISA and Western Blot, respectively. The antiserum titer was about 1:400 000, and the specificity of the antiserum was confirmed by Western Blot. The prepared antiserum can provide experimental materials for the functional identification of HcDMC1 protein.

Key words: Halostachys caspica, HcDMC1, prokaryotic expression, fusion protein, antiserum

中图分类号:

Q949.745.1

张冀, 张丽丽, 张贝贝, 张富春. 盐穗木HcDMC1的原核表达和抗血清的制备[J]. 植物研究, 2018, 38(3): 399-405.

ZHANG Ji, ZHANG Li-Li, ZHANG Bei-Bei, ZHANG Fu-Chun. Prokaryotic Expression of Halostachys caspica HcDMC1 and Preparation of Its Antiserum[J]. Bulletin of Botanical Research, 2018, 38(3): 399-405.

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盐穗木HcDMC1的原核表达和抗血清的制备

Prokaryotic Expression of Halostachys caspica HcDMC1 and Preparation of Its Antiserum

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