Optimized DNA sequence, recombinant plasmid, strain, recombinant protein, mycoplasma gallisepticum antibody colloidal gold detection test paper and detection card
A technology of DNA sequence and Mycoplasma gallisepticum, which is applied in the directions of measuring devices, instruments, scientific instruments, etc., can solve problems such as colloidal gold test strips that have not disclosed Mycoplasma gallisepticum antibody, and achieve the effect of improving the detection effect.
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Embodiment 1
[0082] 1. Preparation of MG-VlhA1.2 recombinant protein
[0083] 1.1 Acquisition and optimization of Mycoplasma gallisepticum VlhA1.2 gene and construction of recombinant plasmid pGEX-KG-VlhA1.2
[0084] The method for obtaining and optimizing the sequence of the Mycoplasma gallisepticum VlhA1.2 gene involved in the present invention and the method for constructing the recombinant plasmid pGEX-KG-VlhA1.2 Expression [J], Chinese Journal of Veterinary Medicine, 2007.
[0085] The specific implementation method is as follows: obtain the VlhA1.2 gene sequence according to the pMGA1.2 sequence included in GenBank (accession number: AF275312). Since the UGA codon in Mycoplasma encodes tryptophan, and UGA is a stop codon in Escherichia coli, in order to achieve high-efficiency expression of the target gene in Escherichia coli, TGA was optimized to TGG; The gene was optimized according to the preference of Escherichia coli, and an EcoR I (GAATTC) restriction site was added to the 5'...
Embodiment 2
[0139] Evaluation of the stability of the colloidal gold detection test paper for mycoplasma fowl septicemia antibody of the present invention
[0140] The vacuum-packaged test papers of the present invention placed at 4°C and 37°C respectively were taken out on the 7th, 16th, 35th, 54th, 70th, and 90th day, and tested after the standard positive serum of Mycoplasma gallisepticum was diluted with pure water. . The results are shown in Table 2. The test paper of the present invention has a longer shelf life at 4° C., and the shelf life is at least 90 days.
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