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

Construction method of REV virus infectious cloning for expressing green fluorescent envelope fusion protein

A technology of REV virus and fusion protein, which is applied in the field of construction of REV virus infectious clones, can solve problems such as inappropriate insertion sites, and achieve the effect of improving efficiency and simplifying the construction process

Active Publication Date: 2016-01-27
YANGZHOU UNIV
View PDF3 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Scholars at home and abroad also tried to insert the green fluorescent protein gene into the REV genome to rescue the REV tracer virus expressing green fluorescent protein, but due to the inappropriate insertion site selected, the REV virus expressing the green fluorescent envelope fusion protein has not been successfully obtained so far.

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
  • Construction method of REV virus infectious cloning for expressing green fluorescent envelope fusion protein
  • Construction method of REV virus infectious cloning for expressing green fluorescent envelope fusion protein
  • Construction method of REV virus infectious cloning for expressing green fluorescent envelope fusion protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0028] 1) Design and amplify REV infectious clone linearization vector and EGFP gene fragment primers: amplify the upstream primer of REV infectious clone linearization vector located at position 6112-6141 of the proviral genome of SNV strain; amplify the linearization of LTR containing REV virus The downstream primer of the vector is located at positions 6082-6111 of the proviral genome of the SNV strain; the upstream primer for amplifying the EGFP gene has 15 EGFP gene-specific bases at the 5 end in addition to the 18 EGFP gene-specific bases starting from the EGFP gene ATG Bases that are reverse-complementary to the downstream primers of the linearized vector for amplifying the infectious clone of REV; the downstream primers for amplifying the EGFP gene contain 15 conserved and amplified containing Bases complementary to primers upstream of the REV infectious clone linearized vector. See attached table 1 for specific primer sequences, which were synthesized by LifeTechnolog...

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 of REV virus infectious cloning for expressing green fluorescent envelope fusion protein. According to the construction method, green fluorescent EGFP gene segments amplified by the specific PCR primer are inserted into the linearized N end of Env genes of an REV viral genome, the commercial recombinase Exnase TM II is adopted for directly carrying out rapid recombinant cloning in vitro without the enzyme-cut and link up reaction, then the green fluorescent EGFP gene segments after the rapid recombinant cloning are transferred to the escherichia coli to be subjected to plasmid amplification and identification, and positive clone transfection DF1 cells are rescued to obtain REV viruses for expressing the green fluorescent envelope fusion protein.

Description

technical field [0001] The invention relates to a method for constructing an infectious clone of REV virus expressing green fluorescent envelope fusion protein. This invention can not only obtain the REV tracer virus expressing the green fluorescent envelope fusion protein, which fills the gap at home and abroad; it provides materials for the study of REV virus replication in the body, tissue tropism and its pathogenic mechanism; Viral vectors offer the possibility to express foreign genes. Background technique [0002] Avian reticuloendotheliosis virus (REV) mainly causes a group of syndromes characterized by reticuloendothelial cell hyperplasia, including acute reticulosis, dwarf syndrome and chronic neoplastic hyperplasia of lymphoid tissue and other tissues disease. REV-infected chickens tend to grow stunted, and the elimination rate and mortality increase; and cause immunosuppression of infected chickens, interfere with the immune effect of other poultry disease vacci...

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): C12N15/10C12N15/63
Inventor 叶建强周晓祥秦爱建邵红霞
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