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 |
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Authors: | 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 |
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Institution: | 1. Interuniversity Centre for Genomics and Gene Technology, Department of Biotechnology, University of Kerala, Trivandrum, Kerala, India;2. Regional Medical Research Centre (ICMR), Port Blair, Andaman and Nicobar Islands, India;3. Madurai Kamaraj University, Madurai, Tamil Nadu, India |
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Abstract: | 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 kinetics and protein expression compared to the strain harbouring the cytosolic protein gene. |
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Keywords: | Corresponding author at: Department of Botany Institute of Science Banaras Hindu University Varanasi UP India |
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