Production of Recombinant Human Interferon alpha 2b (rhIFN?-2b) in genetically engineered strain of Bacillus subtilis

Authors

  • Saira Ashfaq Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories Complex Lahore, Pakistan.
  • Sania Mazhar Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories Complex Lahore, Pakistan.
  • Naaz Abbas Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories Complex Lahore, Pakistan.
  • Ramsha Essa Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories Complex Lahore, Pakistan.
  • Bakhtawar Bukhari Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories Complex Lahore, Pakistan.
  • Khizra Ismail Yasar Saleem
  • Yasar Saleem Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories Complex Lahore, Pakistan.
  • Quratulain Syed Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories Complex Lahore, Pakistan.
  • Sana Riaz Section Animal Physiology and Neurobiology, Department of Biology, KU Leuven, Belgium.

DOI:

https://doi.org/10.52700/jmmg.v5i1.158

Keywords:

IFNa-2b, Bacillus subtilis, Cyt1 promoter, KO7, NBG medium, SDS-PAGE.

Abstract

Interferon alpha 2b (IFN?-2b) is a critical therapeutic protein with immunomodulatory properties, widely used in the treatment of various diseases, including viral infections and cancer. This study presents a comprehensive methodology for the cloning and characterization of the IFN?-2b gene, including the identification of point mutations, followed by expression in Bacillus subtilis. The research begins with gene isolation, primer design, RNA extraction, cDNA synthesis, and PCR amplification. Cloning into a T/A vector and subsequent sequencing reveal point mutations within the gene. The transformed construct is expressed in Bacillus subtilis, and the resulting protein is analyzed using SDS-PAGE. The identification of point mutations and successful protein expression in Bacillus subtilis. This study provides valuable insights into the IFN?-2b gene and its potential applications.

Published

2024-04-30