Codon optimization of CI protein coding sequence of potato virus Y and establish-ment of eukaryotic expression system and preparation of monoclonal antibody
The nucleic acid sequence encoding the CI protein of potato virus Y(PVY)had multiple regions similar to prokaryotic promoter elements.It was possible to translate toxic proteins in prokaryotic cells,so it was difficult to construct vectors.According to the codon bias of prokaryotes,the CI sequence was modified without changing the amino acid sequence.The expression vector containing CI open reading frame(ORF)was success-fully constructed,and CI protein was successfully expressed through the eukaryotic cell-free protein expression system.Six cell lines were prepared by immunizing Balb/c mice with purified protein using hybridoma technolo-gy.Indirect ELISA and western blot showed that the prepared CI monoclonal antibody 4B7_2D6(IgGl)had high sensitivity and specificity.The successful expression of CI protein provides a prerequisite for the purification of CI protein and the subsequent study of the structure and function of CI protein,which is of great significance for further exploring the interaction mechanism between CI and plant proteins.
CI proteincodon biaseucaryotic expressionmonoclonal antibody
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