Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for identifying active region of ATP5B (adenosine triphosphate 5B) gene promoter

A technology of active regions and promoters, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc.

Inactive Publication Date: 2018-10-16
GANSU AGRI UNIV
View PDF1 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the transcriptional regulation mechanism of bovine ATP5B gene

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for identifying active region of ATP5B (adenosine triphosphate 5B) gene promoter
  • Method for identifying active region of ATP5B (adenosine triphosphate 5B) gene promoter
  • Method for identifying active region of ATP5B (adenosine triphosphate 5B) gene promoter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Analysis of luciferase activity of ATP5B gene promoter

[0037] 1. Design of promoter primers

[0038] According to the 5′UTR sequence of the ATP5B gene in GeneBank, the transcription start site identified by 5′-RACE is +1, and gene-specific primers were designed by Clone Manage software:

[0039] The upstream primer is Forward Primer F1: 5'-CAGTCTTTCTCCTTTATTCCTCTGCTA-3'

[0040] The downstream primer is Reverse Primer R1: 5'-TCTGCTTATCACTCTGGGCG-3'.

[0041] Bovine genomic DNA was used as a template for PCR amplification, and the target fragment was 1897bp in total from the promoter sequence -1972 to -76. Add 1.2μL (100ng) DNA template, 0.4μL KOD-Plus-Ver 2.0 (1U / μL), 2μL 10×PCR Buffer, 2μL dNTP (2mmol / L), 0.8μL MgSO to a 200μL PCR tube 4 (25mol / L), 0.6μL ForwardPrimer F1 (10μM), 0.6μL Reverse Primer R1 (10μM), 12.4μL ddH 2O, a total of 20 μL, vortexed and centrifuged, placed in a PCR instrument for reaction, the reaction conditions are as follows: pre-denatura...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a method for identifying the active region of an ATP5B (adenosine triphosphate 5B) gene promoter. The method comprises the following steps: (1) cloning an ATP5B gene promoter sequence; (2) constructing an ATP5B gene 5' end promoter series deletion fragment luciferase reporter vector; and (3) performing cell transfection and luciferase activity analysis on the ATP5B gene 5' end promoter series deletion fragment luciferase reporter vector in the step (2) respectively, and indicating the active area of the ATP5B gene promoter is an area between the two adjacent deletion fragments if the activity of the promoter between the adjacent two deletion fragments is obviously changed. The active area of the ATP5B gene promoter is identified by a luciferase activity analysis method, a regulating and control component for regulating and controlling the transcriptional activity of the ATP5B gene promoter is further identified, and a MyoD and GATA transcription factor binding site is determined to be the regulating and control component for regulating and controlling the transcriptional activity of the ATP5B gene promoter.

Description

technical field [0001] The invention relates to a method for identifying the active region of the ATP5B gene promoter, and also relates to a method for identifying the regulatory elements regulating the transcriptional activity of the ATP5B gene promoter. Background technique [0002] ATP5B (Adenosine triphosphate 5B, Adenosine triphosphate 5B) is the protein encoded by the β subunit of ATP synthase. For the whole enzyme, the β subunit is the catalytic subunit. In eukaryotic cells, it catalyzes the synthesis of ATP The rate-limiting reaction is an important protein in the oxidative phosphate pathway in the process of cellular energy metabolism. Studies have found that the α and β subunits of ATP synthase exist on the plasma membrane surface of 3T-3L1 preadipocytes, and the expression of ATP synthase in the plasma membrane increases significantly during the process of adipocyte transformation. Arakaki et al. used different small molecule inhibitors of ATP synthase and specif...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6897C12N15/85
CPCC12N15/85C12Q1/6897
Inventor 赵志东昝林森胡江刘秀
Owner GANSU AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com