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

Human membrana cofactor protein mini-gene pmcp constructing method

A technology of synergistic factors and construction methods, applied in the field of pathogenic biology, which can solve the problems of unsuccessful development of transgenic animal models, etc.

Inactive Publication Date: 2008-05-07
YANGZHOU UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, transgenic animal models for many human-specific pathogenic infectious diseases are still not successfully developed

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
  • Human membrana cofactor protein mini-gene pmcp constructing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0018] Construction process of the present invention as shown in Figure 1, is divided into the following steps:

[0019] 1. Construction of receptor MCP minigene

[0020] (1) Utilize PCR technology, take human blood cell genomic DNA as the template to amplify the promoter of MCP:

[0021] The amplified upstream and downstream primer sequences are as follows:

[0022] 5′-GC ACGCGT GGTAACTGCCAGAAATTCTTTA-3' and 5'-GTACAGCAGCAACA CCATGG-3', the 5' end of the upstream primer was introduced into the Mlu I restriction site.

[0023] 50μl PCR amplification system includes: 5μl 10× buffer, 2mmol / L Mg 2+ , 200μmol / LdNTPs, 200nmol / L forward and reverse primers, 2μg human blood cell genomic DNA, 2.5U LA TagDNA polymerase. The PCR program for 30 cycles is 94°C / 50s (the first cycle is 5min), 57°C / 40s, 72°C / 2min (the last cycle is 10min). The amplified products were analyzed by electrophoresis on 1% agarose gel and recovered and purified.

[0024] (2) Using PCR technology, using pSG5-...

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 construction method for human membrane cofactor protein mini gene, belonging to pathogenic biology field, which is characterized that a human blood cell genome DNA is selected as a template, and a promoter of the human membrane cofactor protein gene is cloned with a PCR method; a human membrane cofactor protein mini gene cDNA is amplified with pSG5-MCP template; then the MCP promoter is spliced with the cDNA with the PCR overlapping method; a CMV promoter of the pcDNA3.1 is replaced by the obtained DNA fragment, and the human membrane cofactor mini gene pMCP is constructed. A transgenic mouse built by the invention can change the situation that the ideal animal model is scarce in prior study of human specific pathogen infectious disease, and break through the limits of infectious pathogenesis of corresponding pathogen, vaccine evaluation, drug evaluation and other aspects.

Description

technical field [0001] The present invention relates to the field of pathogenic biology. Background technique [0002] Pathogens can be divided into three categories according to different pathogenic objects, most of which can infect humans as well as animals, and are zoonotic pathogens, such as rabies virus, bird flu virus, etc.; the second category is only Pathogens that infect animals, such as Newcastle disease virus; the third category is pathogens that only infect humans. There are not many types of such pathogens, but they may cause great damage to humans, such as Neisseria gonorrhoeae and Neisseria meningitidis , measles virus, human herpes virus-6, HIV, etc., the losses brought to human society can be said to be well known. Human beings are the only hosts of these pathogens, and there is a lack of effective animal models for their pathogenic mechanisms and control strategies, so they are greatly limited. Some researchers use cell models or tissue culture models to ...

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
IPC IPC(8): C12N15/12C12N15/09
Inventor 李国才季明春龚卫娟王劲松朱立天潘兴元王晓红焦红梅
Owner YANGZHOU 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