Polypeptide for marking cytoskeleton protein and eukaryotic expression carrier thereof

A eukaryotic expression vector and cytoskeleton technology, applied in the biological field, can solve the problems of high price and achieve the effects of cost reduction, clear operation steps and mature process operation

Inactive Publication Date: 2010-11-10
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Companies that produce this fluorescently labeled phalloidin include Invitrogen and Sigma, but they are expensive

Method used

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  • Polypeptide for marking cytoskeleton protein and eukaryotic expression carrier thereof
  • Polypeptide for marking cytoskeleton protein and eukaryotic expression carrier thereof
  • Polypeptide for marking cytoskeleton protein and eukaryotic expression carrier thereof

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Embodiment Construction

[0044] The present invention provides polypeptides that can bind to F-actin and are applied to the labeling of cytoskeletal proteins. The polypeptides include JN27, JN30 and TatJN27; the polypeptide JN27 or JN30 binds a fluorescent substance to the intracellular The microfilament has a static indicator effect; after the polypeptide TatJN27 is combined with a fluorescent substance, it has a targeted indicator effect on the microfilament of living cells, and can observe the dynamic change of the microfilament;

[0045] Furthermore, the present invention constructs a fusion expression vector of JN27, JN30 polypeptide and green fluorescent protein; transfects them into cells to observe the dynamic changes of microfilaments.

[0046] The present invention will be described in detail below, which is an explanation rather than a limitation of the present invention.

[0047] 1. Screening of peptides that bind to F-actin

[0048] The JN molecule is a molecule specifically expressed by oligoden...

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Abstract

The invention discloses a polypeptide for marking cytoskeleton protein and an eukaryotic expression carrier thereof. After the polypeptide is transferred into a cell or the polypeptide is expressed in the cell through the eukaryotic expression carrier, the polypeptide can be joined with a microfilament, and the polypeptide marked with fluorescence can be used for marking the cytoskeleton protein. The polypeptide is of JN27, JN30 or TatJN27; and the eukaryotic expression carrier is of pEGFP-C2-JN27 or pEGFP-C2-JN30. The JN27, JN30 and TatJN27 polypeptide being marked with fluorescence is induced into the cell, after pEGFP-C2-JN27 and pEGFP-C2-JN30 eukaryotes express plasmid transferred cell, the pEGFP-C2-JN27 and pEGFP-C2-JN30 eukaryotes can be joined with micro-filament protein in the skelemin in the cell so as to realize the fluorescence marking of skelemin in the cell.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and relates to the labeling of cytoskeleton proteins, in particular to a polypeptide and eukaryotic expression vector for marking cytoskeleton proteins. Background technique [0002] Movement is one of the most distinctive features of life forms. As a unit of life, the various physiological activities of cells are closely related to cell movement, such as the change and maintenance of cell morphology, intracellular material transport, cell division, endocytosis, and exocytosis. The cytoskeleton system that exists in cells (especially eukaryotic cells) not only supports the structure of living cells, determines the shape of cells, and gives them strength, but is also the material basis for the spatial arrangement of various organelles in cells. The cytoskeleton includes three types of protein fibers, namely microfilament, microtubule and intermediate filaments (IF). [0003] Microfilaments are fibrous stru...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K1/13C12N15/62C12N15/79G01N21/64
Inventor 张斌王涛药立波
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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