Engineering E. coli for the biosynthesis of nanoparticles

• Main Author: Koh, Zhong Ren
• Other Authors: Lim Sierin
• Format: Final Year Project
• Language: English
• Published: 2016
• Subjects: DRNTU::Science::Chemistry::Analytical chemistry::Proteins; DRNTU::Engineering::Bioengineering; DRNTU::Science::Biological sciences::Microbiology::Bacteria; DRNTU::Science::Biological sciences::Genetics
• Online Access: http://hdl.handle.net/10356/68471
• Institution: Nanyang Technological University

With the potential of Archaeoglobus fulgidus ferritin to serve as an MRI contrast agent, the study sought to optimize ferritin and feoB production using arabinose induction and to optimize in vivo iron loading in strains of engineered E. coli. The plasmid pBbE8k was primarily used as a vector for ferritin and feoB genes. Arabinose induction for feoB was roughly optimized at 0.1 mM. However, narrower optimization and repeatability studies are highly recommended in future. The AfFtn gene in pBbE8k failed to be expressed although many efforts were taken to resolve this issue throughout the course of the year. Nevertheless, the expression of IPTG-induced AfFtn in pET-11a was achieved and the results of the Bradford Assay, Iron Colorimetric Assay, and Dynamic Light Scattering for this strain serve as a point of reference for the pBbE8k strain. The re-cloning of RBS and ferritin gene onto pBbE8k by varied methods is highly recommended in order to make progress in this ongoing effort.