Pardis Saeedi, Elham Behzadi, Rohallah Kazemi, Mohammad Javad Rezaei, Jafar Amani,
Volume 9, Issue 36 (9-2019)
Abstract
Escherichia coli is the first bacteria on which cloning and protein expression are carried out. The purpose of this study was to summarize the numerous articles published in the field of protein expression in Escherichia coli. In studies, researchers in a new project require the production of pure protein, which at the theory level is easy to obtain a recombinant protein, but there are virtually many problems, including low growth of host, formation of inclusion, protein not being active, and there is even no protein production in the exprimental scale. To enhance the expression of the protein, factors such as suitable host, vector design, transcriptional settings, selective markers, and reluctant labels are effective. There are a range of fusion proteins that can effect on accurate folding, solubility, and proteolytic resistance. Also the ability to target produced proteins to cytoplasm, periplasm, membranes and the culture medium, and some techniques grant the ability of post translational modifications in prokaryotic cells. Therefore, the key to the successful expression of recombinant proteins in E. coli is a combination of expert manipulation of the components of a broad genetic apparatus.
Mohammadali Yaghobi Moghaddam, Alireza Saeedinia, Afshin Samimi Nemati, Mohammadjavad Dehghan,
Volume 11, Issue 43 (7-2021)
Abstract
Aim and Background: With the advent of cloning technology and recombinant DNA, it became possible to produce drugs and vaccines that were not possible in the past. Recombinant protein occupies a large part of the biopharmaceutical industry, one of the most important of which is vaccination.
Material & Methods: In this study, with the aim of obtaining a protein candidate for cholera vaccine, first cloning of a plasmid designed for the bacterial host E.coli DE3 Rossetagami was transformed, then expression was induced using IPTG and after protein expression was examined by SDS-PAGE gel electrophoresis, and then the protein was purified by nickel-sepharose (NI-NTA) chromatography column.
Result:The results show that the purified protein has a molecular weight of 31 kDa and the amount of purified protein is 1 mg.
Conclusion:Based on previous studies as well as the results of this study, the resulting protein can be considered as a suitable candidate for the cholera vaccine.
