Echinococcus granulosus 3-hydroxyacyl-coa dehydrogenase protein and its application
A technique for Echinococcus granulosus, hydroxyacyl, in the field of immunology
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Embodiment 1
[0037]
[0039] The EGR-03347 gene (as shown in SEQ ID NO: 2) was amplified by PCR with the protoscoria cDNA as a template.
[0041] Reaction program: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30s, annealing at 62°C for 30s, extension at 72°C for 30s, a total of 35 cycles
[0044] The results show: the size of the target fragment and the expected result are consistent with 822bp (see Figure 1).
[0049] (2) After 20min, take out and place in a 42° C. water bath for heat shock for 90s, and immediately transfer to an ice bath on ice for 5min.
[0055] Reaction program: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30s, annealing at 62°C for 30s, extension at 72°C for 30s, a total of 35 cycles
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[0064] After mixing the above components, connect overnight at 4°C. The ligation product was transferred into DH5α competent cells, and the transformation steps were the same as
[0074] 3. the next day, the thalline was repeatedly ...
Embodiment 2
[0099] The experimental dogs were immunized with an equal volume of 200 μg protein content and a final concentration of 2 mg / mL QuilA solution.
[0101] The EGR-03347 group, the PBS group, and the QuilA experimental dog were injected with immune substances at two subcutaneous points on the neck respectively at 200 μg / . waived
[0107] (5) Display: add 50 μL of TMB chromogenic solution to each well, and incubate at 37° C. for 15 min.
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PUM
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