Abstract:
Objective The polyclonal antibody against MiPDCD6 protein in the esophageal glands of Meloidogyne incognita was prepared to study the pathogenic mechanism of the root-knot disease on plants transmitted by the nematode.
Method The functional fragment of MiPDCD6 was amplified by PCR to construct recombinant plasmid pET-32a-MiPDCD6 and transform it into Escherichia coli BL21 cells for MiPDCD6 fusion protein induction and expression. Polyclonal antibodies were prepared by immunizing male New Zealand white rabbits with purified MiPDCD6 expression protein. Titer and purification of the obtained antibody were verified using ELISA and SDS-PAGE techniques.
Result Under IPTG concentration of 1.0 mmol·L−1 at 37℃with constant rotation of 150 r·min−1 for 5 h, MiPDCD6 transformed in the E. coli BL21 was clearly expressed. The secured polyclonal antibody was highly specific with a high titer of approximately 1∶50000 as shown by ELISA and SDS-PAGE.
Conclusion The prokaryotic expression conditions of MiPDCD6 were determined. The polyclonal antibody obtained had a high titer and specification against MiPDCD6 and was considered adequate for studying the pathogenesis of the root knot disease on plants infected through M. incognita.
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