Cloning and expression of fusion protein Glutathion s transferase-streptavidin on Bacillus subtilis spores and evaluation of its specific binding with biotinylated antibody cetuximab

  • Bùi Thu Thủy
  • Phan Hương Trang
  • Nguyễn Trần Hiếu
  • Trần Thị Mỹ
  • Phan Tuấn Nghĩa
  • Trần Thị Vân Anh

Abstract

We cloned fusion gene (cotB-gsi-streptavidin), encoding three sequential proteins including membrane spore coat protein CotB, Glutathion s Transferase (GST), and streptavidin, into the chromosome of Bacillus subtilis PY79 to create recombinant strain named SA2. Expression of streptavidin on the outer coat of SA2 spores were confirmed by immunofluoresence using specific anti-streptavidin rabbit IgG and anti-rabbit IgG labeled Alexa546. The specific bindings of biotinylated cetuximab on spores of Bacillus subtilis SAI (expressing CotB-streptavidin) and on Bacillus subtilis SA2 were detected by Western Immunoblotting. The data indicates that binding of biotinylated cetuximab on SA2 spores was slightly 1.4 fold higher than that on SAl spores, suggesting limited role of GST in enhancing the exposure of streptavidin for interaction with biotinylated cetuximab.

điểm /   đánh giá
Published
2017-09-20
Section
Articles