Construction method of lentivirus expression plasmid carrying FGF-1 gene

A technology of FGF-1 and expression plasmid, which can be applied in the direction of virus, genetic engineering, plant genetic improvement, etc., can solve the problem of low mitogenic ability of recombinant FGF1 protein

Inactive Publication Date: 2019-04-12
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows researchers to easily transfer genes into different types of cancerous tissue by introducing an enzyme called rhenium (R) that breaks down DNA molecules inside tumors without harming healthy ones nearby. These modifications help scientists better understand how these changes affect their own body's ability to fight off disease effectively.

Problems solved by technology

The technical problem addressed in this patents relates to developing better treatments for diabetic nephropathy without affecting ongoing therapy or reducing their risk of death due to cardiovascular events associated with high levels of glucose accumulation over time.

Method used

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Comparison scheme
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Embodiment Construction

[0017] Embodiment of the present invention: construction method of lentiviral expression plasmid carrying FGF-1 gene

[0018] (1) FGF-1 gene amplification

[0019] 1. FGF-1 gene PCR amplification

[0020] According to the FGF-1 gene sequence (Gene ID: 2246) in NCBI, use the software LSPrimer (http: / / ccsipb.lnu.edu.cn / primer / ) to design the specific primers FGF-1-F and FGF of the FGF-1 gene -1-R, add the enzyme cutting site Not I and Avr II, the introduced enzyme cutting site Not I sequence is detailed in the sequence table SEQ ID 5, the introduced enzyme cutting site AvrII sequence is detailed in the sequence table SEQ ID 6, the For the sequence of the specific primer FGF-1-F, see SEQ ID 1 in the sequence listing, and for the sequence of FGF-1-R, see SEQ ID 2 in the sequence listing.

[0021] Use the pEmcherry-N1-FGF-1 plasmid as a template for PCR amplification. The PCR reaction system is:

[0022] PCR reaction program: Denaturation at 94°C for 4 minutes, renaturation at 5...

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Abstract

The present invention discloses a construction method for a lentivirus expression plasmid carrying a FGF-1 gene. The FGF-1 gene is inserted into Not I and Avr II multiple cloning sites of a pLenti-Mcherry-Puro vector, MDA-MB-453 cells are transfected with a constructed pLenti-FGF-1-Emcherry-Puro recombinant plasmid, and a cell line stably transfected with the FGF-1 gene is obtained by puromycin screening and identification. The recombinant plasmid contains red fluorescence protein-containing mcherry, which is convenient for observing positive cells under a fluorescence microscope; and the recombinant plasmid carries a puromycin resistance gene tag and an ampicillin resistance gene tag, and ampicillin or the puromycin can be added during prokaryotic expression or eukaryotic expression for screening. The plasmid lays a solid foundation for further studies of biological characteristics and functions of the FGF-1 gene and research and development of anti-tumor drugs.

Description

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Claims

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Application Information

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Owner GUIZHOU UNIV
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