Combined protein for mycobacterium tuberculosis detection and detection reagent
A technology of Mycobacterium tuberculosis and combined protein, which is applied in the field of medical immune detection, can solve the problems of difficult promotion, high price, long detection period and the like
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Embodiment 1
[0039]The preparation method of detection reagent for tuberculosis mycobacterium is as follows:
[0040]1), the synthetic gene fragment of the first combined protein of the present invention and the second composite protein of the present invention is cut from the cloned carrier, and XhoI is cut, cloned to the expression vector PET28B, in E. coli BL21 (DE3) The expression was incorporated to the OD600 was 0.6, and IPTG was added to the final concentration of 0.1 M induced expression, and analyzed by gel electrophoresis and WesternBlotting;
[0041]2), use gel (Pharmacia CHELANG SEPHROSE FAST FLOW), and the expression protein is purified by metallic chromatography;
[0042]3), using standard immunization procedural immunized rabbits, preparation of rabbits (IgM, IGA) by standard immunization programs (purchased from Tiangen Biocuro).
[0043]4), prepared by citric acid reducing method; ie, 0.01% aqueous solution of aqueous solution of aqueous solution of aqueous solution is heated, and 1% aqueou...
Embodiment 2
[0053]The preparation method of detection reagent for tuberculosis mycobacterium is as follows:
[0054]1), the synthetic gene fragment of the first combined protein of the present invention and the second composite protein of the present invention is cut from the cloned carrier, and XhoI is cut, cloned to the expression vector PET28B, in E. coli BL21 (DE3) The expression was incorporated to the OD600 was 0.6, and IPTG was added to the final concentration of 0.1 M induced expression, and analyzed by gel electrophoresis and WesternBlotting;
[0055]2), use gel (Pharmacia CHELANG SEPHROSE FAST FLOW), and the expression protein is purified by metallic chromatography;
[0056]3), using standard immunization procedural immunized rabbits, preparation of rabbits (IgM, IGA) by standard immunization programs (purchased from Tiangen Biocuro).
[0057]4), prepared by citric acid reducing method; ie, 0.01% aqueous solution of aqueous solution of aqueous solution of aqueous solution is heated, and 1% aqueou...
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