Prokaryotic Expression of Halostachys caspica HcDMC1 and Preparation of Its Antiserum

doi:10.7525/j.issn.1673-5102.2018.03.012

Abstract

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

CLC Number:

Q949.745.1

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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