Cloning, Expressing, and Purifying the IsoCitrate Lyase-1 Enzyme in Mycobacterium Tuberculosis
Advisor(s)
Dr. Angela Ahrendt, Illinois Mathematics and Science Academy
Location
Room B133
Start Date
26-4-2019 9:45 AM
End Date
26-4-2019 10:00 AM
Abstract
In this study, we chose to target the Isocitrate Lyase 1(ICL-1) gene as a possible target for the treatment of tuberculosis. We took this approach because ICL-1 allows the Mycobacterium tuberculosis bacteria to skip some of the steps in the KREBS cycle, which gives it the ability to survive in low-oxygen environments. The ICL-1 gene found in M. tuberculosis was amplified using PCR to prepare an expression system for testing drugs targeting the ICL-1 protein. In order to do this, primers were designed to clone the gene and give it restriction sites, which allowed us to insert the ICL-1 gene insert into a vector, and finally into E. coli cells. Since this work is still in progress, an expression vector containing ICL-1 was acquired and transformed into BL21 DE3. Protein was subsequently expressed and purified.
Cloning, Expressing, and Purifying the IsoCitrate Lyase-1 Enzyme in Mycobacterium Tuberculosis
Room B133
In this study, we chose to target the Isocitrate Lyase 1(ICL-1) gene as a possible target for the treatment of tuberculosis. We took this approach because ICL-1 allows the Mycobacterium tuberculosis bacteria to skip some of the steps in the KREBS cycle, which gives it the ability to survive in low-oxygen environments. The ICL-1 gene found in M. tuberculosis was amplified using PCR to prepare an expression system for testing drugs targeting the ICL-1 protein. In order to do this, primers were designed to clone the gene and give it restriction sites, which allowed us to insert the ICL-1 gene insert into a vector, and finally into E. coli cells. Since this work is still in progress, an expression vector containing ICL-1 was acquired and transformed into BL21 DE3. Protein was subsequently expressed and purified.