Perpustakaan Digital ITB

Despite the varying important roles of membrane proteins such as ion transport, transmembrane signaling, secondary metabolites synthesis, and many more, the expression of heterologous membrane protein in prokaryotes remains a challenge. Formation of inclusion body, competition of expression between the heterologous and the essential protein, inactivated protein, and death of the host cells due to toxicity are still commonly encountered. One of the strategies to overcome these challenges is by fusing Mistic (Membrane-Integrating Sequence for Translation of Integral Membrane Protein Construct) with the membrane protein. Mistic has been proven in assisting the heterologous membrane protein expression in Escherichia coli. However, it has not been implemented yet for heterologous membrane protein expression in Bacillus subtilis. In this research, the mistic – green fluorescent protein construct (mstX-gfp) in pDR111 plasmid was transformed to B. subtilis and its expression were observed using a fluorescence and confocal laser scanning microscope. The construct has been successfully transformed and the expression showed the emission of green fluorescence from GFP. However, the localization of GFP in the membrane cannot be concluded yet and requires a further study.