Akhtar Khadija-Tul Kubra and Idah Sithole-Niang*
Department of Biochemistry, University of Zimbabwe, Mt. Pleasant, Harare, Zimbabwe
*Corresponding Author: Idah Sithole-Niang, Department of Biochemistry, University of Zimbabwe, Mt. Pleasant, Harare, Zimbabwe.
Received: February 05, 2018; Published: March 07, 2018
Citation: Akhtar Khadija-Tul Kubra and Idah Sithole-Niang. “Development of a Cloning Vector for Biologicals and Therapeutics”. Acta Scientific Pharmaceutical Sciences 2.4 (2018).
Molecular cloning is a technique used to manipulate an organism’s genome using biotechnology. This provides a way of overcoming barriers to gene transfer between species. The aim of this study was to remove a gene from one organism and transfer it to another so that the gene is expressed in the new host. A Nuclear inclusion (NIa) protease gene coding for an autocatalytic protease obtained from Cowpea aphid-borne mosaic virus (CABMV) was fused with the green fluorescence protein gene in a pUC57 plasmid. The fusion was then cloned into a pUC18 expression vector via the SmaI site. Escherichia coli (E. coli) NM522 cells were transformed by this recombinant vector. Protein expression was induced using Isopropyl β-D-thiogalactopyranoside (IPTG). The protein sample was analysed using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis which showed that the protein was successfully expressed and cleaved out of the fusion vector system by the protease.
Keywords: Gene Cloning; Autocatalytic Protease; Cowpea Aphid-Borne Mosaic Virus; Protein Expression; Therapeutic
Copyright: © 2018 Akhtar Khadija-Tul Kubra and Idah Sithole-Niang. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.