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

Method for producing human serum albumin by using animal mammary gland and expression vector thereof

A technology of human serum albumin and carrier, which is applied in the field of transgenic technology, can solve the problems of low expression level of human serum albumin, and achieve the effect of low cost and increased profit

Active Publication Date: 2014-07-02
SHANGHAI INST OF MEDICAL GENETICS SHANGHAI CHILDRENS HOSPITAL
View PDF3 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a method for producing human serum albumin with a high expression level of human serum albumin in the existing method for producing human serum albumin by using the mammary gland of transgenic animals. Method for producing human serum albumin by animal mammary gland and expression vector used therein

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
  • Method for producing human serum albumin by using animal mammary gland and expression vector thereof
  • Method for producing human serum albumin by using animal mammary gland and expression vector thereof
  • Method for producing human serum albumin by using animal mammary gland and expression vector thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1. Fragment synthesis

[0044] 1) Amplification of hAlb cDNA fragment.

[0045] Use upstream primer Alb-F: 5'-CTGTCAACCCCACACGCCTT-3' (SEQ ID NO.1), downstream primer Alb-R: 5'-TT TTCATTTTTCTTTCT-3' (SEQ ID NO.2), human peripheral blood cell cDNA was used as a template for PCR amplification. The reaction system is as follows:

[0046]

[0047] A total of 4 reaction tubes were made, and the reaction conditions were 94°C for 5min; (94°C for 45sec, 56°C for 45sec, 72°C for 3min) × 32 cycles; 72°C for 10min. A hAlb cDNA fragment of about 2.5 kb was amplified. After mixing the reaction solution, 5 μL was sent to Shanghai Boshang Biotechnology Co., Ltd. for sequencing. The remaining product was subjected to agarose electrophoresis, and the gel was cut to recover the 2.5 kb hAlb cDNA fragment with a kit. Then the sequence obtained by sequencing was compared with the sequence NM 000477.5 in the Genebank database, and it was found to be completely consistent.

[0048] 2)...

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 discloses a method for producing human serum albumin by using mammary glands of transgenic animals and an expression vector thereof. The method provided by the invention comprises the steps of: using a vector to transfer a human serum albumin gene expression cassette to a genome of a mammal, so as to obtain a transgenic animal; and acquiring human serum albumin from the milk of the transgenic animal. The human serum albumin gene expression cassette from 5' to 3' includes a promoter, a human serum albumin gene and a terminator in order, also includes one or two selected from a 3' flanking sequence M2 of the human serum albumin gene and a 3' flanking sequence B3 of a beta-casein gene, wherein M2 or B3 are located after the terminator. According to the method provided by the invention, the concentration of human serum albumin in animal milk can at least reach 4.6-6.7 g / L, which can fully meet the commercial utilization level. The method isolates human serum albumin from milk of transgenic cow, so as to provide a low-cost and high-yield method for human serum albumin production.

Description

technical field [0001] The present invention relates to transgenic technology, in particular to a method for producing human serum albumin using mammary glands of transgenic animals with high expression level of human serum albumin and the expression vector used therein. Background technique [0002] Human serum albumin is the most abundant protein in plasma, which plays an important role in maintaining plasma colloid osmotic pressure, nutrition, and material transport in the body, and is widely used in the treatment of burns, shock, malnutrition, and chronic wasting diseases. At present, almost all the albumin needed clinically is the plasma of blood donors. However, due to the difficulty of insufficient blood sources, especially the risk of infection with hepatitis HIV1 virus and HIV when using blood donors' plasma, mass production of safe albumin has always been a difficult and urgent goal to achieve. [0003] "The promoter of goat beta casein gene guides the specific ex...

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): C12N15/85C12N15/14C07K14/765
Inventor 曾溢滔曾凡一蔡勤方彧聃黄淑帧
Owner SHANGHAI INST OF MEDICAL GENETICS SHANGHAI CHILDRENS HOSPITAL
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