Inactivation of chromosomal genes of the choline bacterial microcompartment through transformation of Escherichia coli 536

Bacterial microcompartments (BMCs) are proteinaceous organelle structures inside a bacterium that allow enzymatic activity to occur within a controlled space. Several different types of BMCs have been identified including microcompartments that facilitate the breakdown of choline into products that can be used by the cell. While research has revealed general information about the protein shell of BMCs, not much is known about the specific genes that are involved in the choline microcompartment. This project was conducted to determine the effects inactivation of genes implicated in the choline microcompartment would have on Escherichia coli 536 and its ability to breakdown choline into trimethylamine (TMA). Using an amended version of the Wanner and Datsenko protocol for transformation which involves using PCR products to inactivate chromosomal genes, transformants were tested for their ability to breakdown choline. As of now, the study is still ongoing, and due to high numbers of false positives the antibiotic kanamycin used in transformation is being changed to chloramphenicol. Identifying if the transformants can breakdown choline has yet to be determined for all genes. However, the results of this study can provide information regarding the relationship between TMA, E. coli 536, and cardiovascular and urinary tract associated diseases.