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BCG vaccine strain of recombined interferon and its preparation process

A technology of recombinant interferon and BCG, which is applied in the field of genetic engineering, can solve the problems of wide application, high cost and repeated perfusion, etc., and achieve the effect of avoiding high cost, increasing immune effect and reducing dosage

Active Publication Date: 2008-03-12
天津市泌尿外科研究所 +5
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  • Abstract
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Problems solved by technology

However, the high cost of this solution and the need for repeated perfusion have affected its further widespread application.

Method used

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  • BCG vaccine strain of recombined interferon and its preparation process

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Embodiment Construction

[0027] The present invention will be described in detail below in conjunction with the accompanying drawings and implementation.

[0028] 1. Materials and methods

[0029] Plasmid: The vector plasmid pMAO-4 and the plasmid containing hIFN-α-2B cDNA were provided by Professor YI LUO of Iowa University, USA.

[0030] 2. Method

[0031] (1) PCR reaction of hIFN-α-2B cDNA fragment:

[0032] Upstream primer: CAAGggatccTGTGATCTGCCTCAAACCCACAG, 50 μmol / l;

[0033] Downstream primer: GCCGgaattcTCATTCCTTACTTAAACTTTCTT, 50 μmol / l.

[0034] (2) BamHI and EcoRI digestion of pMAO-4 plasmid and hIFN-α-2BcDNA PCR product The pMAO-4 plasmid and hIFN-α-2BcDNA were ligated overnight in a 16°C water bath with T4 DNA ligase.

[0035] (3) Transformation of plasmids:

[0036] Add 100 μl competent bacteria DH-5α to each tube, add 1 μl phIFN-α-2B plasmid, and inoculate the transformed samples on LB solid medium containing 30 μg / ml kanamycin. Select positive colonies. Many scattered pale yellow...

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Abstract

The invention discloses a recombinant interferon--BCG Vaccine bacterial strain and its preparing method, transferring the composed pshuttle phIFN-alpha-2B into the BCG to compose Rbcg-hIFN-alpha-2B bacterial strain. Because it can secrete the interferon IFN-alpha-2B, it can reduce the usage of BCG as compared with that of the wild BCG on the condition of reaching the same and even increased immune effect, thus reducing toxic and side effect; and it can secrete cell factors at the properest time and part. Therefore, it lays the foundation of application to curing cystic tumors.

Description

technical field [0001] The invention relates to genetic engineering technology, in particular to a recombinant interferon BCG strain and a preparation method thereof. Background technique [0002] Bladder cancer is the most common malignant tumor of the urinary system in my country. According to statistics, about 75% of the cases are superficial bladder cancer, and 95% of them are high-risk superficial tumors. If no drug treatment is given after surgery for high-risk superficial tumors, the short-term recurrence rate is 60-90%. Therefore, the prevention of postoperative recurrence of bladder tumors and the treatment of postoperative residual tumors and carcinoma in situ are major clinical problems to be solved urgently. [0003] Bacillus Calmette-Guerin (BCG) is a biological immunomodulator formed by the highly virulent Mycobacterium bovis. It is widely used to prevent tuberculosis all over the world. It has been proved to have a high degree of safety and few serious compli...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/20C07H21/00C12Q1/68A61K39/04A61P35/00C12R1/32
Inventor 韩瑞发刘春雨姚智韩育植马腾骧
Owner 天津市泌尿外科研究所
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