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

Gene of expressing regeneration enhancement factor of recombined human liver in full lenght, and method for preparing regeneration enhancement factor of recombined human liver in full length, and use

A technique of enhancing factors and liver regeneration, applied in biochemical equipment and methods, botany equipment and methods, genetic engineering, etc., can solve the problem of lack of liver specificity

Inactive Publication Date: 2008-05-28
北京地坛医院
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unlikely to be key regulators of liver regeneration due to the lack of liver-specific actions of most factors

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
  • Gene of expressing regeneration enhancement factor of recombined human liver in full lenght, and method for preparing regeneration enhancement factor of recombined human liver in full length, and use
  • Gene of expressing regeneration enhancement factor of recombined human liver in full lenght, and method for preparing regeneration enhancement factor of recombined human liver in full length, and use
  • Gene of expressing regeneration enhancement factor of recombined human liver in full lenght, and method for preparing regeneration enhancement factor of recombined human liver in full length, and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Preparation process for expression of recombinant full-length human liver regeneration enhancer factor Pichia pastoris

[0023] 1. Preparation of DNA Select the amino acid sequence of the full-length human liver regeneration enhancer factor CAB87993 from Genebank, such as SEQ ID NO.1, with an isoelectric point of 7.2 and a molecular weight of 23kDa, and analyze the corresponding DNA according to the codon diagram Nucleotide sequence, and redesign the DNA sequence of the full-length human liver regeneration enhancer suitable for Pichia expression according to the principle of preferred codons in Pichia pastoris, and submit the obtained sequence table to "Shanghai Boya Biotechnology Company" for DNA sequence Synthesis, the synthesized DNA sequence was submitted to "Shanghai Shenyou Biotechnology Co., Ltd." for DNA sequence sequencing and identification, and the DNA nucleotide sequence as described in SEQ ID NO.2 was obtained;

[0024] 2. Ligate the DNA nucleotid...

Embodiment 2

[0030] Example 2 Recombinant full-length human enhancer of liver regeneration promotes DNA synthesis of liver cancer cell line HepG2 in vitro

[0031]1. HepG2 cell culture: HepG2 cells were grown in high-glucose DMEM+10% fetal bovine serum, 37°C, 5% CO 2 nourish. After the cells grow to the logarithmic phase, trypsinize and adjust the cell density to 4×10 5 / ml, inoculate 100 μl per well in a 96-well culture plate, and continue culturing for 6 h.

[0032] 2. Adding samples: Add the samples to be tested in the 96-well culture plate, with a total of 3 concentrations (20ng / ml, 50ng / ml, 100ng / ml), set 3 duplicate wells for each concentration, and continue to cultivate for 24h. Join 2.0×10 4 Q 3 H-TdR, continue to culture for 3h.

[0033] 3. Detection: Use a cell harvester to collect the cells on a filter paper sheet, and count them with a liquid scintillation counter (see Table 1).

[0034] Table 1 In vitro promotion of DNA synthesis of liver cancer cell line HepG2

[0035]...

Embodiment 3

[0037] Example 3 Recombinant full-length human enhancer of liver regeneration promotes DNA synthesis in rat liver cells after partial hepatectomy in vivo

[0038] 1. Establishment of rat partial hepatectomy model: Rats were fed in single cages for 1 week, fasted overnight before the experiment, anesthetized by intraperitoneal injection of pentobarbital sodium (40mg / kg) before the operation, and 750mL / L after depilation of the upper abdomen. Disinfect the skin with alcohol, make a midline incision in the upper abdomen, and expose the liver. After the ligaments of the left and middle lobes of the liver are fully dissociated according to the Hoggin method, the roots of each liver lobe are firmly ligated, and the left and middle lobes of the liver are removed quickly at the distal end of the ligature. That is equivalent to 2 / 3 liver resection. The remaining liver lobe was preserved, and the abdominal incision was sutured.

[0039] 2. Dosing: 1 ml (0.1 mg / ml) of recombinant full-l...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to View More

Abstract

A gene for expressing recombinant full-length augmenter of liver regeneration (ALR) is disclosed. A process for preparing said recombinant full-length ALR includes such steps as artificially synthesizing its gene sequence according to its amino acid sequence, inserting it into plasmid Ppic9k, TRANSFERRING IT IN Pichia yeast GS115, yeast expression and protein purifying to obtain high-purity product. It can be used for treating chronic hepatitis, serious hepatitis and hepatocirrhosis.

Description

technical field [0001] The invention belongs to the production method and application field of bioengineering products, and specifically relates to a DNA nucleotide sequence expressing recombinant full-length human liver regeneration enhancing factor through Pichia pastoris and its expression vector, and using the above DNA expression vector to prepare the A method for recombining the full-length human enhancer of liver regeneration factor and a pharmaceutical use of the recombinant full-length human enhancer of liver regeneration factor. Background technique [0002] The liver is one of the organs with the most powerful regeneration ability in the human body. The regeneration of the liver is of great significance in the pathogenesis and treatment of various liver diseases such as acute / chronic viral hepatitis and severe hepatitis. The regulatory mechanism has been continuously studied, and dozens of regulatory factors have been found, such as hepatocyte growth factor (HGF),...

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 Patents(China)
IPC IPC(8): C12N15/12C12N15/63C12P21/02C12N15/81A61K38/17A61P1/16
Inventor 成军洪源
Owner 北京地坛医院
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