Preparation method for polyclonal antibody of specific recognition protein PRM2 and application of preparation method

A polyclonal antibody and protein recognition technology, applied in the direction of serum immunoglobulin, anti-animal/human immunoglobulin, material inspection products, etc., can solve problems such as being in the blank stage, and achieve easy access, high expression yield, Physical fitness effect

Inactive Publication Date: 2019-12-03
NANJING MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] However, the relevant research on the preparation method of protein PRM2 polyclonal antibody in the prior art is still in a blank stage

Method used

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  • Preparation method for polyclonal antibody of specific recognition protein PRM2 and application of preparation method
  • Preparation method for polyclonal antibody of specific recognition protein PRM2 and application of preparation method
  • Preparation method for polyclonal antibody of specific recognition protein PRM2 and application of preparation method

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Experimental program
Comparison scheme
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Embodiment

[0026] The polyclonal antibody is a mouse IgG2a subtype polyclonal antibody.

[0027] A method for preparing a polyclonal antibody that specifically recognizes protein PRM2, comprising the steps of:

[0028] (1) Obtain the PRM2 protein fragment; select the target fragment by antigenic epitope analysis for recombinant expression of protein PRM2, the gene sequence of the target fragment is shown in SEQ ID No.1 in the sequence table, and its corresponding amino acid sequence is shown in sequence As shown in SEQ ID No.4 in the list, BamH1, NotI restriction sites and homology arms were introduced into the upstream and downstream primers respectively;

[0029] An upstream primer, the specific sequence of which is shown in SEQ ID No.2 in the sequence listing;

[0030] A downstream primer, the specific sequence of which is shown in SEQ ID No.3 in the sequence listing;

[0031] (2) extract human normal kidney tissue mRNA, and use it as a template to perform reverse transcription PCR,...

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Abstract

The invention relates to the technical field of biological science and detection reagents, in particular to a preparation method for a polyclonal antibody of specific recognition protein PRM2 and application of the preparation method. The preparation method comprises the following steps that (1) antigen epitope analysis of the protein PRM2 is conducted, specifically, a target fragment is selectedthrough antigen epitope analysis as recombinant expression of the protein PRM2, a gene sequence of the target fragment is shown in the SEQ ID NO.1 in a sequence list, and BamH1 and Not I digestion sites are correspondingly introduced into an upstream primer and a downstream primer. According to the preparation method, the preparation of the polyclonal antibody of the PRM2 is achieved.

Description

technical field [0001] The invention relates to the technical fields of biological science and detection reagents. Background technique [0002] The problem of infertility has always been an important topic in the research of human reproduction. Now 15% of couples in the world suffer from this problem, about half of which are caused by the man. The study of its mechanism is essential for finding solutions to infertility. [0003] During the haploid stage of vertebrate spermatogenesis, protamine replaces histones in sperm chromatin and becomes the major DNA-binding protein in the sperm nucleus. With the help of protamine, the DNA double strand undergoes a series of coiling, folding, and compression to form high-density chromosomes, whose volume is less than 5% of the somatic nucleus. Most mammals have only one type of protamine (protamine 1); however, some species, including humans and mice, have two types of protamine: protamine 1 and protamine 2 (PRM2, protamine 2). Stu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K16/06G01N33/68
CPCC07K16/06C07K16/18G01N33/689G01N2333/47
Inventor 刘明兮刘思钰吴佳艳王莹
Owner NANJING MEDICAL UNIV
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