Recombinant calmette-Guerin bacillus vaccine for secretion of human interferon-alphaza and its constructing method

A technology for recombinant BCG and its construction method, which is applied in the field of recombinant BCG that secretes human interferon-α2a and its construction, to achieve the effects of improving drug efficacy, reducing the burden of medical expenses, and reducing medical expenses

Inactive Publication Date: 2005-12-21
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

However, the research on improving the curative effect of bladder tumor by constructing recombinant BCG capable of secreting IFN-α2a by means of genetic engineering technology has not been reported yet.

Method used

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  • Recombinant calmette-Guerin bacillus vaccine for secretion of human interferon-alphaza and its constructing method
  • Recombinant calmette-Guerin bacillus vaccine for secretion of human interferon-alphaza and its constructing method
  • Recombinant calmette-Guerin bacillus vaccine for secretion of human interferon-alphaza and its constructing method

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Embodiment 1

[0032] Step 1 Construction of BCG Shuttle Expression Vector

[0033] 1. Synthesis of amplification primers

[0034]According to the gene sequences of Ag85B signal peptide and human IFN-α2a, the main secretion antigens of BCG, which are already available in GeneBank, the amplification primers for Ag85B signal peptide and IFN-α2a were designed, respectively.

[0035] Ag85B sense: 5′-GGATCCATGACAGACGTGAG-3′

[0036] antisense: 5′-CTGCCGTGCCGATCATCAAT-3′

[0037] IFN-α2a sense: 5′-AGCGAATTCTGTGATCTGCCTCAAACCCA-3′

[0038] antisense: 5′-AGCGTCGACTCATTCCTTACTTCTTAAACTTTCTTGC-3′

[0039] Sequencing primer on pMV261 vector: 5′-GACAACTTGAGCCGTCCGTC-3′

[0040] 2. Load the Ag85B signal peptide gene into the pMV261 vector

[0041] Synthesize the signal peptide gene and clone it into pUCm-T vector

[0042] Ligation (TAKARA) system: pUCm-T vector 1μl

[0043] Signal peptide fragment (about 50ng / μl) 4μl

[0044] Solμtion I 5μl ...

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Abstract

This invention involves reorganization BCG vaccine for secretion of people interferon- alpha2a, and structuring method. Utilize gene engineering to clone BCG vaccine Ag85B signal peptide part and the genes of people IFN - alpha 2a playing a secreting role to pMV261 to receive the BCG vaccine shuttling expression carrier pMSIFN - alpha 2a. Adopt electric boring technology to lead pMSIFN - alpha 2a into BCG to recombinate rBGGIFN - alpha2a. Depending on rBGGIFN - alpha 2a duplicating in BCG and the secretion function of the signal peptide, can secrete IFN - alpha2a efficiently.

Description

technical field [0001] The present invention relates to a recombinant BCG that secretes human interferon-α2a and its construction method, in particular to a recombinant BCG containing the main secretion antigen Ag85B signal peptide fragment of BCG and interferon-α2a (IFN-A 2A) and its construction method The obtained recombinant bacillus Calmette-Guerin rBCGIFN-α2a is used for preventing and treating superficial bladder tumors, and belongs to the technical field of genetic engineering. Background technique [0002] Since Morales first reported the application of Bacillus Calmette-Guerin (BCG) bladder infusion in the treatment of superficial bladder tumors in 1976, BCG as an effective immune adjuvant has achieved good results in the prevention and treatment of bladder tumors. One of the most effective adjuvant treatments for tumors. Although BCG has good efficacy in the prevention and treatment of superficial bladder tumors, there are still many problems to be solved. The f...

Claims

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Application Information

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IPC IPC(8): C12N15/09
Inventor 刘海涛夏术阶孙晓文
Owner SHANGHAI JIAO TONG UNIV
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