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

Muscle enhancer sequence and application thereof

A nucleotide sequence and promoter technology, applied in the field of muscle enhancers, to achieve the effect of improving promoter activity

Active Publication Date: 2013-02-06
龚纮毅
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still room for improvement in the activity of the zebrafish mylz2 promoter in other fish species and species than zebrafish

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
  • Muscle enhancer sequence and application thereof
  • Muscle enhancer sequence and application thereof
  • Muscle enhancer sequence and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1: Construction of expression vectors for novel fluorescent protein genes TcFP-11 and TcFP-13 of Taiwan coral

[0075] In this experiment, primers were designed using the cDNA sequences of Taiwan coral fluorescent protein genes TcFP-13 and TcFP-11. The PCR reaction conditions were as follows: denaturation at 94°C for 2 minutes; denaturation at 94°C for 2 minutes; denaturation at 94°C for 30 seconds; annealing and renaturation at 55°C for 30 seconds seconds; synthesis at 68°C for 1 minute; denaturation, annealing and extension steps repeated for 35 cycles; final extension reaction at 72°C for 10 minutes, and finally stored at 4°C. After colloid extraction, the reaction product was ligated with the cloning vector (pGEM-T-easy vector), and then transferred into recipient cells; after a small amount of culture, the cloning vector was extracted for sequencing, and the sequence data was compared with the original sequence data. Compare, and extract a medium amount of ...

Embodiment 2

[0076] Example 2: Molecular selection of zebrafish ckmb gene regulatory sequence and construction of expression vector

[0077] In this experiment, the genome browser Ensembl Genome Browse (http: / / www.ensembl.org) was used to search the zebrafish ckmb gene, and the binding position of transcription factors related to muscle growth was predicted from the start codon (ATG) to the 5' end. Use the online database program (Genomatix, www.genomatix.de, MatInspector program) to search and predict, with a reliability of 80% as the selection index; select the following three fragments as the clipping range and design primers accordingly Amplify: 2363bp fragment (-1202 / +1161), 1271bp fragment (-1202 / +69) and 1343bp fragment (-182 / +1161) (see sequence position Figure 4 ), set the following reaction conditions: 94°C denaturation for 2 minutes; 94°C denaturation for 30 seconds; 72°C for 10 minutes, and finally stored at 4°C. After colloid extraction, the reaction product was ligated wit...

Embodiment 3

[0078] Example 3: Fluorescence qualitative experiment of zebrafish eggs

[0079] Using the fluorescent expression vector of Example 2, the Tol2 transfer system was used to conduct a fluorescent qualitative experiment on zebrafish eggs, and the expression activities of ckmb-P2363 fragments, ckmb-P1271 fragments and ckmb-P1343 fragments were analyzed.

[0080] Collect zebrafish (Danio rerio) and arrange them in egg trays, mix the microinjection solution (100ng / μL) containing carrier DNA and Tol2 transposon mRNA (100ng / μL) at a ratio of 1:1, and inject (IM-300 Microinjector, Narishige) was injected with a microscope (LEICAEZ4, LEICA). The injection conditions were set at an injection pressure of 7-10psi, and an injection time of 100msec, which was slightly adjusted depending on the actual conditions. The injected solution (in the color of pheno red dye Observation) occupies about 1 / 4 of the egg. The needle is inserted from the embryonic cell at an angle of 45 degrees. The injecti...

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 relates to a muscle enhancer from a zebra fish muscle type creatine kinase b (ckmb) gene and application thereof. The enhancer can improve the activity of a promoter and can be used for preparing transgenic animals and applied to bioreactors.

Description

technical field [0001] The present invention relates to a muscle enhancer from zebrafish muscle creatine kinase b (ckmb) gene and application thereof. Background technique [0002] Gene transfer is a technology widely used in the field of biotechnology today, which is to introduce nucleic acid molecules that can express specific gene products into cells through genetic engineering technology, so as to modify their gene expression, give new performance traits or serve as a means of mass production of specific gene products tool. For example, for economic animals such as cattle, sheep, pigs, fish, and shrimp, it can be used to improve varieties, improve meat quality, increase growth rate, improve disease resistance and adaptability, or increase ornamental value. Taking fish as an example, the consumption pattern of the aquarium industry has gradually changed from traditional breeding and ornamental to artistic, refined and life-oriented. The development and innovative applic...

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): C12N15/113C12N15/63C12N5/10A01K67/033
Inventor 龚纮毅陈鸣泉吴金洌黄士晋
Owner 龚纮毅
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