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Method for culturing staphylokinase transgenic tomato

A staphylokinase and staphylokinase encoding technology, applied in the field of cultivating staphylokinase-transformed tomato, can solve the problems of low staphylokinase production and high price

Inactive Publication Date: 2010-12-15
BEIJING FORESTRY UNIVERSITY
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  • Abstract
  • Description
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Problems solved by technology

[0003] At present, staphylokinase produced by Escherichia coli as a bioreactor has appeared on the domestic market, but the produced staphylokinase has low yield and high price

Method used

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  • Method for culturing staphylokinase transgenic tomato
  • Method for culturing staphylokinase transgenic tomato
  • Method for culturing staphylokinase transgenic tomato

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, the acquisition of transgenic tomato and its detection

[0036] 1. Obtaining genetically modified tomatoes

[0037] 1. Construction of recombinant plant expression vector

[0038] The recombinant expression vector pBSAK containing the staphylokinase SAK gene was constructed by the following method: according to the SAK sequence in the relevant patent (application number: CN200710053340.0), the SAK gene sequence (sequence 2 in the sequence listing) was cloned from Staphylococcus aureus 1-489th nucleotide sequence), the gene encodes the protein of sequence 1 in the sequence table; the staphylokinase SAK gene is cloned into plasmid pBI121 (purchased from Beijing Baierdi Biotechnology Co., Ltd., article number: BIODEE MP-091) Between the XbaI and BamHI restriction sites, the structure of the T-DNA region formed is as follows figure 1 shown.

[0039] 2. Transformation of tomato plants

[0040] (1) Recombinant expression vector pBSAK transforms Agrobacterium...

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Abstract

The invention discloses a method for culturing a staphylokinase (SAK) transgenic tomato, comprising the following steps: (1) building a recombinant plant expression vector containing an SAK encoding gene; and (2) converting a tomato plant into the SAK transgenic tomato by the built recombinant plant expression vector. The method of the invention can help successfully obtain the SAK transgenic tomato plant; and a solusphere method proves that SAK protein expressed by the transgenic plant has thrombolytic activity, thus laying the foundation for large-scale production of SAK by taking the tomato as a bioreactor.

Description

technical field [0001] The invention relates to a method for cultivating transgenic plants, in particular to a method for cultivating transstaphylokinase gene tomato. Background technique [0002] Cardiovascular and cerebrovascular diseases such as cerebral thrombosis and hypertension seriously endanger people's health, and the age of onset is gradually getting younger. The development of drugs for the treatment of cardiovascular diseases is a research hotspot today. Staphylokinase (Staphylokinase, SAK) is a plasminogen activator secreted by Staphyloccocus aureus Rosenbach, first discovered in 1948, it can activate plasminogen in the human body to form fibrin lysozyme, thereby dissolving the thrombus. Compared with other thrombolytic drugs, staphylokinase has the advantages of good thrombolytic effect, strong fibrin selectivity, and no significant coagulation-promoting effect. It is one of the hot spots in the research of new thrombolytic agents. [0003] At present, staph...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/58C12N9/48C12Q1/68C12Q1/34G01N33/53A01H5/00
Inventor 陆海刘頔杜景川梁宏伟陈发菊李凤兰
Owner BEIJING FORESTRY UNIVERSITY
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