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Carrier containing immunoglobulinlg gene, construction method and uses thereof

A technology of immunoglobulin and its construction method, which is applied in the field of vectors containing immunoglobulin genes and their construction, and can solve problems such as the construction of fusion expression vectors for human IgG1 Fc segments that have not been seen yet

Inactive Publication Date: 2009-08-19
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there has been no report on the construction of a fusion expression vector using the human IgG1 Fc segment

Method used

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  • Carrier containing immunoglobulinlg gene, construction method and uses thereof
  • Carrier containing immunoglobulinlg gene, construction method and uses thereof
  • Carrier containing immunoglobulinlg gene, construction method and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Implementation 1 Construction of recombinant vector pBV220 / Fc

[0044] 1. Capture the gene encoding the Fc fragment of the heavy chain constant region of human immunoglobulin IgG1 (Human lmmunoglobulin gamma-1)

[0045] The specific method is as follows:

[0046] According to the reported human IgG1 gene sequence (Genbank accession number is BC006402), design and synthesize specific primers. The Fc fragment caught is located at the 816-1523bp position of the above known sequence. The primers are as follows:

[0047] Upstream primers: ATGGAGCCCAAATCTTGTGACAAAACTC-3'(S EQID No.2)

[0048] EcoR I

[0049] Downstream primers: G CGATCCGCCACCGCC CCGTCGCACTCATTTACCC

[0050] BamH I Xhol I linker (linker)

[0051] G-3' (SEQ ID No.3)

[0052]The upstream primer contains an EcoR I restriction site, and the downstream primer has a gene encoding a 5-peptide Gly-Gly-Gly-Gly-Ser flexible linker (linker). The 5 amino acids encoded by this gene ha...

Embodiment 2

[0070] Example 2 Take the fusion expression of staphylokinase (SA K) as an example to illustrate the application of this vector in prolonging the half-life of recombinant protein drugs in vivo

[0071] Natural staphylokinase (staphylokinase, Sak) is a single-chain protein synthesized by lysogenic phage of Staphylococcus aureus, consisting of 136 amino acids. Staphylokinase itself has no enzymatic activity, it can combine with plasminogen on the surface of thrombus 1:1 to form a complex, and further activate the fibrinolytic system. Because Sak activates plasmin with fibrin specificity and low bleeding tendency, the thrombolytic effect of Sak does not affect the levels of fibrinogen, plasminogen and α2-antiplasmin in plasma, and it is effective for old thrombus And platelet-rich thrombus has a stronger lytic effect than traditional thrombolytic drugs, so Sak is a promising thrombolytic agent.

[0072] 1. The method of cloning SAK into the recombinant expression vector pBV220 / F...

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Abstract

The invention discloses a vector containing immunoglobulin gene, belonging to the field of molecular biology. The nucleotide sequence of the vector is shown in SEQID No.1. The invention also discloses a method for constructing the vector, and the vector can be used for remarkably prolonging the half life period of recombinant protein drug in vivo, thus prolonging the drug efficacy, reducing the times of medicine use of patients and lowering the cost.

Description

technical field [0001] The present invention relates to a carrier, in particular to a carrier containing an immunoglobulin gene and its construction method, and taking the fusion expression of staphylokinase (SAK) as an example, the carrier is used to prolong the half-life of recombinant protein drugs in vivo Applications. Background technique [0002] The use of DNA in vitro recombination technology to obtain the target protein is the basis of protein structure and function and bioengineering medicine research. The continuous deepening of genomics and proteomics research has greatly expanded the research field of new protein drugs. After decades of exploration, the high-efficiency expression technology of foreign genes in prokaryotic cells has become relatively mature, and a large number of recombinant proteins expressed through this method have been used clinically. However, most of the first-generation genetically engineered protein drugs currently used clinically have ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N15/66C12N1/21C12P21/02A61K38/48A61K47/48C12R1/19
Inventor 蔡欣徐东刚邹民吉王金凤王园园
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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