SAPNA BHORIA* AND POOJA MALIK
Centre for Biotechnology, M. D. University, Rohtak-124 001 (Haryana), India
*(e-mail: bhoriasapna@gmail.com; Mobile: 94665 32143)
(Received: January 20, 2023; Accepted: February 24, 2023)
ABSTRACT
The inefficient production and insensitivity of human body to insulin have caused diabetes in a large section of human population worldwide. In this study, CTB was fused with proinsulin gene and cloned downstream of the soybean seed-specific glycinin (Gy1) promoter in binary vectors, pBinGy1red2 with a DsRed2 reporter gene. CTB was conjugated at the N-terminus of the proinsulin gene to prevent degradation and also to facilitate purification and uptake into the gut- associated lymphoid tissue (GALT) via oral administration. The CTB.pro-insulin also contained three furin cleavage sites that made possible for its in vivo processing (outside the pancreas) in most of the mammalian body cells. The cloned binary vectors were transferred into E. coli (for plasmid maintenance) and Agrobacterium tumefaciens (for plant transformation).
Key words : CTB-proinsulin, binary vector, cloning, Agrobacterium, heat shock