%A YU Guo-Wu, QING Yun, HE Shan, HUANG Yu-Bi %T Preparation and application of polyclonal antibody against SSIIb protein from maize %0 Journal Article %D 2022 %J Acta Agronomica Sinica %R 10.3724/SP.J.1006.2022.03010 %P 259-264 %V 48 %N 1 %U {https://zwxb.chinacrops.org/CN/abstract/article_7245.shtml} %8 2022-01-12 %X

Antibody is an important reagent for protein function research. It is difficult to carry out protein function research due to the lack of corresponding antibody. Maize starch synthase IIa and starch synthase IIb (SSIIb) belong to starch synthase, which are mainly responsible for the extension of amylopectin. However, the studies of starch synthase IIb were still in lack. In this study, we immunized New Zealand white rabbits with recombinant GST fusion protein (455-704 aa) at the C-terminal of starch synthase IIb as antigen and prepared polyclonal antibody against SSIIb protein. Western blot showed that the polyclonal antibody of SSIIb could recognize not only SSIIb-C (455-704 aa) antigen, but also SSIIb protein in endosperm of maize at different developmental stages. Western blot analysis using the polyclonal antibody revealed that the expression pattern of SSIIb protein in maize endosperm increased first and then decreased from 10 to 30 days after pollination, and the highest expression level was found at 15 days after pollination. Semi-quantitative RT-PCR results indicated that the transcription level of SSIIb was the highest at 20 days after pollination. These results indicated that the specific polyclonal antibody against SSIIb was successfully prepared, and it could specifically recognize the endogenous SSIIb protein of maize by Western blot, which laid a foundation for further study of the function of SSIIb protein.