DEVELOPMENT OF LOW-COST EXPRESSION SYSTEMS FOR THE PRODUCTION OF T5 EXONUCLEASE

• Grant number: 22/12656-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2022
• End date: April 01, 2024
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Danielle Biscaro Pedrolli
• Grantee: Lucas Moita Paim
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Associated scholarship(s): 23/06171-3 - Optimization of B. subtillis KO7 fermentation using low-cost substrates in high-throughput microscale bioreactors, BE.EP.IC

Abstract

Modern biotechnology largely employs microorganisms genetically modified through theuse of gene cloning. Among all the procedures involved in gene cloning, the ones thatpresent the most difficulties are digestion by restriction enzymes followed by the bindingof DNA molecules. Cloning is commonly performed using traditional methods ofrestriction enzyme digestion and DNA ligation. But there are already more modernmethods like Golden Gate Assembly. This has advantages over the traditional method,such as the use of only one type IIS restriction enzyme and greater efficiency. But it stillpresents the difficulty in cloning sequences that internally contain the restriction sites ofthe few type IIS enzymes available on the market. On the other hand, the GibsonAssembly method, widely used in laboratories around the world, is often inaccessible tolaboratories in Brazil, mainly due to the high cost and long delivery times of importedproducts. In this context, it is proposed the development of expression systems for theonly component of the Gibson Assembly kit that is not produced in Brazil, T5exonuclease. With this, we intend to enable the production of the kit in the nationalterritory at more affordable prices and deadlines. In addition, development will be opensource, allowing the development of the kit for any national laboratory.