猪RNA聚合酶Ⅱ单克隆抗体的制备与应用

doi:10.11843/j.issn.0366-6964.2021.010.009

Abstract

Abstract: The purpose of this study was to prepare monoclonal antibodies against porcine RNA polymerase Ⅱ and make preliminary applications. The bioinformatics method was used to predict the immunogen, after immunogen was chemically synthesized, 5 female Bal b/c mice aged 4-8 weeks were immunized, and cell fusion experiment was performed on the mice that were positive after immunization, spleen cells of these mice were fused with myeloma cells to obtain hybridoma cells that could stably secrete anti-RNA Pol II antibody. The identification result showed that the heavy chain of RNA PolⅡ monoclonal antibody was IgG 2A type, and the light chain was Kappa type. The hybridoma cells were screened and subcloned by the indirect ELISA method, and 8 hybridoma cell lines that stably secreted RNA PolⅡ monoclonal antibody were obtained. It was preliminary applied to chromatin immunoprecipitation (ChIP-seq) technology, and compared with the enrichment of commercial antibodies, the results showed that the monoclonal antibodies obtained in this study were more enriched. The porcine RNA PolⅡ monoclonal antibody obtained in this study can provide a theoretical basis for epigenetic research and provide important biological materials.

Key words: RNAPⅡ, monoclonal antibody, pig, hybridoma cell

CLC Number:

S852.43

TIAN Xiaohuan, LIU Ru, SUN Yan, REN Ruimin, YU Mei, ZHAO Shuhong, CAO Jianhua. Preparation and Application of Monoclonal Antibody of Porcine RNA Polymerase Ⅱ[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(10): 2783-2791.

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Preparation and Application of Monoclonal Antibody of Porcine RNA Polymerase Ⅱ

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