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

Recombinant expression vector for specifically and highly expressing SIA in muscle-derived cells and application of recombinant expression vector

An expression vector, high-expression technology, applied in applications, vectors, nucleic acid vectors, etc., can solve problems such as high rigidity, influence on insulin activity, and lengthy C-peptide structure, and achieve the effect of avoiding side effects

Active Publication Date: 2021-08-06
SICHUAN UNIV +1
View PDF7 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The C-peptide structure is long and rigid, which affects the activity of insulin. The activity of proinsulin without cutting out the C-peptide is only 5% of that of mature insulin

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
  • Recombinant expression vector for specifically and highly expressing SIA in muscle-derived cells and application of recombinant expression vector
  • Recombinant expression vector for specifically and highly expressing SIA in muscle-derived cells and application of recombinant expression vector
  • Recombinant expression vector for specifically and highly expressing SIA in muscle-derived cells and application of recombinant expression vector

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041]Search the mouse insulin gene (NCBIID: NM_008386.4) in NCBI, download the gene sequence and analyze the nucleic acid sequence of signal peptide, insulin B chain, insulin A chain, insulin C peptide and other parts, and the sequence of C peptide Deleted, replaced with a flexible peptide (GGGGS) 3 Nucleic acid sequence, and enzyme cutting sites are added at both ends of the entire DNA chain. The designed sequence was analyzed by SWISS-MODEL for structure prediction and visualized by VMD. Then entrust Qingke Company to synthesize the DNA sequence and clone it into the pUC57 plasmid, and perform sequencing inspection to ensure that the synthetic sequence is completely accurate. SIA sequence is shown as SEQ ID NO.2.

[0042] Using a flexible peptide (GGGGS)3 to replace the relatively lengthy and rigid C-peptide to connect insulin A and B chains, considering that the length of the chain between insulin B29 and A1 has a certain impact on its activity, its structure is predicte...

Embodiment 2

[0097] SIA plasmid in vivo experiments

[0098] C57BL / 6J male mice were purchased at the age of 6 weeks, adaptively fed for 1 week, and randomly divided into groups: Blank group (normal growth), Control group (modeling, no treatment), Mock group (modeling, injection of pEMS plasmid), Exp1 group (modeling, pCMV-SIA injection) and Exp2 group (modeling, pEMS-SIA injection), 8 mice in each group.

[0099] Establishment of type 1 diabetes mouse model: Weigh the body weight of the mice, add citrate buffer solution to streptozotocin (STZ) and mix well, inject intraperitoneally at a dose of 60 mg / kg, and complete the injection in about 15 minutes. After injection, fast for 2 hours. 1 time a day, continuous injection for 3 days. 72 hours after the injection, the blood of the mice was collected by the tail-cutting method, and the blood glucose was detected with a Roche blood glucose meter and blood glucose test strips. If the blood glucose was greater than 16.7mmol / L and stable for 14...

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 belongs to the technical field of biological medicine, and particularly relates to a recombinant expression vector for specifically and highly expressing SIA in muscle-derived cells and application of the recombinant expression vector. A recombinant expression vector promoter is PEMS, and the nucleotide sequence of the recombinant expression vector promoter is shown as SEQ ID NO.1; and the nucleotide sequence for coding the SIA is as shown in SEQ ID NO. 2. A flexible peptide (GGGGS) 3 is used for replacing a C peptide to modify proinsulin, a high-activity single-chain insulin analogue SIA is obtained, an enhanced muscle specific promoter PEMS is used for regulating and controlling the expression of the SIA, specific efficient expression of the high-activity single-chain insulin analogue SIA in skeletal muscle cells is achieved, the high-activity single-chain insulin analogue SIA is secreted out of cells to enter a blood circulation system, the effects of effectively reducing blood sugar can be achieved, and meanwhile, unnecessary side effects caused by ectopic expression can also be avoided. The recombinant expression vector provided by the invention can be used for preparing medicines for treating type I diabetes mellitus.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a recombinant expression vector specifically and highly expressing SIA in muscle-derived cells and an application thereof. Background technique [0002] Diabetes is a type of disease in which the body's metabolism of glucose is disturbed, resulting in high blood sugar levels in the body. It is greatly affected by genetic and environmental factors. With the development and progress of society and economy, people's lifestyle and eating habits have undergone great changes, leading to a gradual increase in the number of diabetic patients worldwide. Diabetes is usually divided into type 1 and type 2 diabetes. Type 1 diabetes is an autoimmune disease that usually develops in adolescence. Its pathogenesis is mainly that the components of pancreatic beta cells are misrecognized as antigens by the immune system and destroyed, resulting in an absolute lack of insulin in th...

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): C12N15/85C12N15/17C12N15/66A61K38/28A61P3/10
CPCC12N15/85C12N15/66C07K14/62A61K38/28A61P3/10C12N2800/107Y02A50/30
Inventor 王刚邓璐谢丽芳卢宛翎李彩侠刘明
Owner SICHUAN 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