Expression of Leptospira membrane proteins Signal Peptidase (SP) and Leptospira Endostatin like A (Len A) in BL-21(DE3) is toxic to the host cells

Publication date: Available online 2 February 2018 Source:Journal of Genetic Engineering and Biotechnology Author(s): Padikara K. Satheeshkumar, Prasannan V. Anu, Mohmed I. Junaida, Madathiparambil G. Madanan, Tennison Jebasingh, Ananthakrishnan J. Nair, Gangaprasad A. Nair, Govinda Pillai M. Nair, Perumana R. Sudhakaran Heterologous expression of Integral Membrane Proteins (IMPs) is reported to be toxic to the host system in many studies. Even though there are reports on various concerns like transformation efficiency, growth properties, protein toxicity, inefficient expression and protein degradation in IMP overexpression, no studies so far addressed these issues in a comprehensive way. In the present study, two transmembrane proteins of the pathogen Leptospira interrogans, namely Signal peptidase (SP), and Leptospira Endostatin like A (Len-A) were taken along with a cytosolic protein Hydrolase (HYD) to assess the differences in transformation efficiency, protein toxicity, and protein stability when over expressed in Escherichia coli (E. coli). Bioinformatics analysis to predict the transmembrane localization indicated that both SP and Len are targeted to the membrane. The three proteins were expressed in full length in the E. coli expression strain, BL 21 (DE3). Significant changes were observed for the strains transformed with IMP genes under the parameters analysed such as, the transformation efficiency, survival of colonies on IPTG-plate, culture growth kinet...
Source: Journal of Genetic Engineering and Biotechnology - Category: Biotechnology Source Type: research