Construction and validation of more efficient plasmids for the production of recombinant proteins in Escherichia coli

• Grant number: 21/02958-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): June 01, 2021
• Effective date (End): November 30, 2022
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Flavio Henrique da Silva
• Grantee: Thais Fernanda Carlos
• Host Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil
• Associated research grant: 19/26070-1 - Modulation of acquired enamel pellicle to control dental mineral loss: unravelling mechanisms to make therapies possible, AP.TEM

Abstract

Structural and functional studies of proteins require considerable amounts of them, and by using recombinant production systems it is possible to circumvent problems caused by the small amount of purified protein from its natural source. The heterologous expression of proteins with biotechnological interest brings numerous benefits to medicine, pharmaceutical, food, agricultural and chemical fields. However, the yield does not always meet industry demand. In this context, the objective of this work is to develop more efficient plasmids to produce recombinant proteins, altering codons in the 5' fusion from the plasmid pET28, resulting in an N-terminal alteration of the expressed protein, to increase the production yield. As model proteins will be used a phytocystatin from sugarcane, which is already produced and characterized in our laboratory, and one from Maqui, a berry native to South America. Thus, we intend to provide more efficient plasmids in the production of recombinant proteins, and consequently, to obtain a more efficient system for the production of the aforementioned proteins, which have great biotechnological potential. (AU)