Establishment of Prokaryotic Expression and Indirect ELISA Detection Method for Brucella Bp26 Protein
In order to express the Bp26 protein of Brucella in prokaryotic expression,the Bp26 gene fragment was amplified using the Brucella M5-90 vaccine strain.The pET28a-XXA-HRV 3C-Bp26 prokaryotic expression vector was cloned and constructed.The recombinant protein of XXA-HRV 3C-Bp26 was obtained through induction of expression and Ni-NTA affinity purification.Then,the unlabeled Bp26 protein was obtained by PreScission Protease digestion.The results of SDS-PAGE and Western Blot experiments confirmed the successful expression of the XXA-HRV 3C-Bp26 recombinant protein,which was soluble and obtained unlabeled Bp26 protein through enzymatic digestion.Using the Bp26 protein as the diagnostic antigen,a preliminary indirect ELISA detection method for Brucella was established.256 sheep serum samples were tested using a commercial Brucella ELISA detection kit,and the accuracy rate was 93.75%,proving that the established indirect ELISA detection method for Brucella can be used as a clinical detection method for Brucellosis.
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