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Kenaf mitochondrial protein COX3 antigen polypeptide and method for preparing polyclonal antibody and application

A technology of polyclonal antibody and antigen polypeptide, applied in the field of biological protein molecules, can solve the problems of studying the differences in the expression of mitochondrial COX3 gene, and achieve the effect of high sensitivity and strong specificity

Active Publication Date: 2019-05-24
GUANGXI UNIV
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Problems solved by technology

[0004] The present invention proposes a kenaf mitochondrial protein COX3 antigen polypeptide and a method and application for preparing a polyclonal antibody, which solves the problem of studying the expression difference of the mitochondrial COX3 gene in different cytoplasms of kenaf

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  • Kenaf mitochondrial protein COX3 antigen polypeptide and method for preparing polyclonal antibody and application

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Embodiment Construction

[0021] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0022] The process of preparation of kenaf COX3 protein antibody mainly includes the following aspects:

[0023] (1) Kenaf COX3 protein sequence analysis and peptide design and synthesis

[0024] Kenaf according to GenBank (accession number: MF163174 cox3 Design specific primers for amplification of the gDNA sequence cox3 The CDS region sequence of COX3 was analyzed by ORFfinder, and the COX3 protein sequence contained 280 amino acid residues. It was found that its molecular weight was 31.60...

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Abstract

The invention relates to the technical field of biological protein molecules, in particular to a kenaf mitochondrial protein COX3 antigen polypeptide and a method for preparing a polyclonal antibody and an application. An amino acid sequence of the antigen polypeptide is shown in SEQ ID NO: 2. The method includes coupling the antigen polypeptide with carrier protein KLH to obtain polypeptide-KLH coupling composite protein; mixing with a Freund's complete adjuvant with equal volume, emulsifying, and immunizing animals; collecting animal blood serum, collecting animal blood by carotid artery bloodletting when the titer reaches more than 1: 20000, and preparing antibody serum; and purifying the antibody serum by affinity chromatography to obtain the polyclonal antibody of the kenaf mitochondrial protein COX3. By synthesizing the kenaf COX3 polyclonal antibody, the expression difference of the kenaf COX3 protein in different cytoplasm of kenaf is detected, so as to determine the importantinfluence of COX3 on the development of kenaf anthers.

Description

technical field [0001] The invention relates to the technical field of bioprotein molecules, in particular to a method and application for preparing a polyclonal antibody from a kenaf mitochondrial protein COX3 antigen polypeptide. Background technique [0002] Complete mitochondrial gene transcripts are an important basis for studying plant mitochondrial gene expression. CR-RT-PCR is an important method to study the start and end sites of gene transcription, as well as the 5' and 3' transcription splicing patterns of mitochondrial gene transcripts (Kuhn and Binder, 2002; Zhang Dandan, 2016). The presence of multiple transcription initiation sites at the 5' end is a common feature of plant mitochondrial genes. Kuhn et al. found 30 transcription initiation sites in 12 Arabidopsis mitochondrial genes, of which 9 mitochondrial genes contained multiple transcription initiation sites, but only one termination site at the 3' end (Kuhn et al., 2005 ; Forner et al., 2007); Rice HL...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C07K16/16C07K16/06G01N33/68
Inventor 廖小芳赵艳红孔祥军周瑞阳周步进侯文焕唐兴富
Owner GUANGXI UNIV
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