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Gene cloning, expression and characterization of ORF1-C terminal protein of SEN virus D subtype

AIM: To obtain the gene sequence of ORF1-C terminal protein of SENV-D and then to express the molecule, which lays the foundation for further research, diagnosis and therapy applications. METHODS: The SENV-DORF1 C-terminal gene was obtained by PCR from SENV positive serum. After sequencing, the gene was subcloned into pQE30-SEN-D expression vector and transferred into E.coli M15. After IPTG induction, the expressed protein was analyzed by Western blot. RESULTS: The gene sequence of SENV-D ORF1-C terminal protein, as well as the ( M r ) 38×10 3 expressed molecule, was successfully obtained. The expressed protein could be recognized very well by the specific antibody from the patient's serum. CONCLUSION: The SENV-D-ORF1 C-terminal protein is successfully expressed and can be recognized by the specific antibody from the positive serum in Western Blot. It can be used in the detection of SENV-D antibody, which is very useful in monitoring the epidemic situation and detecting the infected patient at early stage.