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Vector pgemt/rhpa and its construction method, and its application in the preparation method of high-efficiency recombinant human plasminogen activator

A carrier and rhpa technology, applied in the direction of recombinant DNA technology, biochemical equipment and methods, using vectors to introduce foreign genetic material, etc., can solve the problems of increasing the risk of clinical application, prone to adverse reactions, and reducing drug safety, etc., to improve Biological activity, low production cost, cost reduction effect

Active Publication Date: 2015-08-26
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a result, production costs increase, resulting in expensive clinical treatment costs
In addition, prokaryotic products have poor compatibility with the human body and are prone to adverse reactions, which also reduces the safety of the drug and increases the risk of clinical application

Method used

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  • Vector pgemt/rhpa and its construction method, and its application in the preparation method of high-efficiency recombinant human plasminogen activator
  • Vector pgemt/rhpa and its construction method, and its application in the preparation method of high-efficiency recombinant human plasminogen activator
  • Vector pgemt/rhpa and its construction method, and its application in the preparation method of high-efficiency recombinant human plasminogen activator

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0032] 1. Carrier construction:

[0033] The technical route of carrier construction is attached figure 1 , the operating steps for constructing the mammary gland characteristic expression component are as follows:

[0034] (1) Materials

[0035] 1.1 Experimental materials and instruments

[0036] DNA amplification instrument (S1000, BIO-RAD, USA), DNA electrophoresis instrument, desktop refrigerated high-speed centrifuge, small desktop high-speed centrifuge (Eppendorf, Germany), constant temperature shaking incubator, redistilled water device, constant temperature water bath shaker, Water-proof electric heating constant temperature incubator, digital gel imager (GIS-1000, Shanghai Tianneng Technology Co., Ltd.), upper dish electronic balance (FA1604, Shanghai Precision Balance), PH detector (HA405-K2, Mettlerto Lido Shanghai Company), ice machine (SIM-F124 Japan Sanyo Company), ultra-clean bench (Suzhou Purification Instrument Equipment Factory), vacuum dryer (Speed ...

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Abstract

The carrier pGEMT / rhPA and its construction method, and its application in the preparation method of high-efficiency recombinant human plasminogen activator belong to the genetic engineering and cell engineering technology in the field of bioengineering. Construct the pGEMT / 5'+3'P vector, and finally construct a pGEMT / rhPA vector with a total of 1068bp at the position 9203-10269, which encodes 356 amino acids. The test proves that the recombinant product expressed has the advantages of high content and high specific activity. It can be used in the development of human thrombolytic drugs.

Description

technical field [0001] The invention belongs to the genetic engineering and cell engineering technology in the field of bioengineering, in particular to the technical field of large-scale recombinant human plasminogen activator (rhPA) production. Background technique [0002] Animal mammary gland bioreactor technology refers to the use of mammalian mammary gland-specific promoter elements to construct transgenic animals, directing the expression of foreign genes in the mammary glands of transgenic animals, and obtaining recombinant proteins from animal milk, which has high commercial value. Value. Rabbits have the advantages of high lactation, short gestation period, and large number of litters per litter. So far, the application of rabbit mammary gland bioreactor to produce recombinant human tissue plasminogen activator has not been reported. [0003] Thrombotic disease is one of the major diseases that endanger human life and health. At present, natural human tissue plas...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/66C12N15/85C12N9/72
Inventor 成勇成姚斌宋绍征葛欣
Owner YANGZHOU UNIV