Agronomy (Jul 2023)

Preparation of Polyclonal Antibody against ZmBT1 Protein and Its Application in Hormone-Regulated Starch Synthesis

  • Lun Liu,
  • Yun Qing,
  • Noman Shoaib,
  • Runze Di,
  • Hanmei Liu,
  • Yangping Li,
  • Yufeng Hu,
  • Yubi Huang,
  • Guowu Yu

DOI
https://doi.org/10.3390/agronomy13071805
Journal volume & issue
Vol. 13, no. 7
p. 1805

Abstract

Read online

In order to investigate the crucial role of ZmBT1 in starch accumulation during maize grain development and analyze the expression and distribution of ZmBT1 in various maize tissues, we prepared a polyclonal antibody. Specifically, we successfully expressed the recombinant plasmid pGEX-6p-ZmBT1-C (382-437aa) and purified Gst-ZmBT1-C as the antigen for antibody preparation. Our results confirmed that the ZmBT1 protein in maize tissues can be specifically recognized by the ZmBT1 antibody. Through Western blotting, we observed that the expression protein of ZmBT1 varied by tissues, with the highest content in the grain and endosperm. Furthermore, we employed a combination of Western blotting and quantitative real-time PCR to show that the expression level of ZmBT1 can be influenced by plant hormones. This finding suggests that ZmBT1 plays a critical role in the accumulation of starch and opens up new avenues for functional studies of this protein.

Keywords