| Abstrakt: |
In this study, the prepared Down syndrome cell adhesion molecule 2 (DSC AM2) polyclonal antibodiesare intended to provide an immunological tool for functional exploration of DSCAM2. This project employs bioinformatics approaches to analyze the DSCAM2 antigenic structure, and constructs DSCAM2828-957 recombinant expression strain by DNA recombination technology. The recombinant DSCAM2828-957 proteinis purified by Ni affinity chromatography and Anti-DSCAM2 polyclonal antibodiesare prepared by immunized mice. Antibody titers are detected by ELISA, antibody specificity is detected by Western Blot, and DSCAM2 localization in Locustamigratoria hemocytes is detected by Wright-Giemsa staining and immunofluorescence. DSCAM2 is knocked down by siRNA, and hemocytes are counted. The results show that the constructed pET30a (+)-DSCAM2828-957/BL21 (DE3) prokaryotic strain can successfully express the recombinant DSCAM2828-957 protein. The immuned mice with DSCAM2828-957 recombinant antigens obtained through Ni chelating affinity chromatography gain polyclonal antibodies with high titer and good specificity. Cell localization analysis shows a high expression of DSCAM2 in basophils of Locustamigratoria. A knockdown of DSCAM2 expression level increases the number of total hemocytes in Locustamigratoria. [ABSTRACT FROM AUTHOR] |
