Immunofluorescence method for locating woody plant pollen tube protein and kit thereof

A technology of woody plants and pollen tubes, applied in the field of plant biology, can solve the difficulties in the study of woody plant pollen tubes, the research of functional proteins has not been reported, etc.

Inactive Publication Date: 2018-07-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows researchers to easily find specific types or parts (polypetide) within wood plant cells that are important functions like photosynthesis. It involves creating an organic mixture containing sugars from sugar beets by cutting off their shells with scissors. These mixtures then reacted with certain chemical agents under controlled conditions such as temperature, pH value, etc., allowing them to bind these molecules together into larger structures called polymer chains. By measuring changes caused when some components have been added during this process, scientists may study how different genotypes affect genetic activity.

Problems solved by technology

This patented describes an approach called transfusional flow cytometry or Western blotting, where specific DNA sequences found within certain types of vegetative material like roots may help determine how well they function properly during various stages of life's history. These techniques require specialized equipment with advanced tools and knowledge about root structure and biological processes. There has been some progress towards understanding this problem because these methods involve cutting off parts of the plant's outer layer - also known as cuff-wall structures – making them hard to analyze further.

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  • Immunofluorescence method for locating woody plant pollen tube protein and kit thereof
  • Immunofluorescence method for locating woody plant pollen tube protein and kit thereof
  • Immunofluorescence method for locating woody plant pollen tube protein and kit thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0050] Example 1 Immunofluorescence method for positioning apple pollen tube protein

[0051] Since the inventor obtained a functional gene in the pollen tube of apple 'Guoguang' by cloning in the early stage, obtained the antigen through prokaryotic expression, and prepared a specific antibody to the rabbit antigen, for the convenience of follow-up research, the pollen of 'Guoguang' was selected as the research object. The process of immunofluorescence method for locating apple pollen tube protein is as follows: germination culture and fixation—enzymatic hydrolysis—blocking—primary antibody immunization—secondary antibody immunization—fluorescence observation. Therefore, experiments were designed from the above steps to determine the location of apple pollen tube protein. Optimal system for immunofluorescence.

[0052] 1. Germination and fixation of apple pollen tubes

[0053] 1. Germination of apple pollen tubes

[0054] Disperse apple pollen on the surface of the liquid m...

Embodiment 2

[0071] Example 2 Localization of exogenously added proteins in apple pollen tubes and co-localization with proteins in pollen tubes

[0072] In order to further verify the feasibility of the immunofluorescence method for locating pollen tube proteins, we took apple pollen as the research object. Since the inventors have previously proved that adding an S nuclease protein to pollen tubes cultured in vitro, the protein can be transmembrane transported to the inside of pollen tubes, and a mouse antigen-specific antibody to the protein has been prepared earlier, so we use Immunofluorescence method for localization of pollen tube proteins was used to detect the localization of exogenously added proteins in pollen tubes. The process is: germination culture and fixation—enzymolysis—blocking—primary antibody immunization—secondary antibody immunization—fluorescence observation. The operation method of this part is consistent with the method in embodiment 1 unless otherwise specified....

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Abstract

The invention provides an immunofluorescence method for locating a woody plant pollen tube protein and a kit thereof. The kit includes a fixing solution, an enzymolysis solution, a blocking solution,a primary antibody and a secondary antibody; the fixing solution contains PEPES, sucrose, Triton X-100, paraformaldehyde, NaCl, KCl, Na2HPO4 and KH2PO4, the enzymolysis solution contains cellulase, macerozyme, NaCl, KCl, Na2HPO4 and KH2PO4, the blocking solution contains bovine serum albumin, NaCl, KCl, Na2HPO4 and KH2PO4, the primary antibody is a target protein antibody, and the secondary antibody is a FITC-labeled goat anti-rabbit secondary antibody. The method adopts an apple pollen tube as a test material, after enzymatic hydrolysis of the wall of the apple pollen tube, immunofluorescentlabeling is performed. The method can directly, efficiently, sensitively and specifically locate the functional protein in the pollen tube, and provides a technical system for visually identifying theexpression of the functional protein in the pollen tube of a woody plant and locating the functional protein in the pollen tube of the woody plant.

Description

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Claims

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Application Information

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Owner CHINA AGRI UNIV
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