European Journal of Biotechnology and Bioscience

ISSN: 2321-9122

Vol. 6, Issue 4 (2018)

Amplification and Cloning of gene encoding of Cell Surface protein (CSP’s) and their Expression in E.coli BL21DE3 strain

Author(s): Arpita Malgi, Rohini B, Kavya R, Priyanka G, Umer Ahmed, Madhavi Mitra, Kausar Parveen, Kelmani Chandrakanth R
Abstract: Escherichia coli also known as E.coli, is a Gram-negative, facultative anaerobic, rod-shaped, coliform bacterium of the genus Escherichia that is commonly found in the lower intestine of warm-blooded organisms (endotherms). Most E.coli strains are harmless, but some serotypes can cause serious food poisoning in their hosts and are occasionally responsible for product recalls due to food contamination. In this study the gene encoding cell surface protein from E.coli was cloned, expressed under the control of T7 promoter and the obtained result showed that Pet19b system was very efficient. the results obtained during this investigation promising conclusion was made as;
  • The CSP gene isolate from E.coli and was incorporated into pET19b vector
  • Transformation was done successfully
  • The clones were analysed for the presence of insert by PCR and they showed that the insert presence
  • The clones were also analysed by SDS-PAGE and one of the clone showed expression of the protein which is 35kdalton.
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