Event Title

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.

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Apr 26th, 9:45 AM Apr 26th, 10:00 AM

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.