1. Rare codons effect on expression of recombinant gene cassette in Escherichia coli BL21(DE3)
- Author
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Hosein Honari, Aghil Esmaeili-Bandboni, and Sadegh Safaei
- Subjects
0301 basic medicine ,Microbiology (medical) ,Genetics ,lcsh:Arctic medicine. Tropical medicine ,Bl21 de3 ,lcsh:RC955-962 ,lcsh:R ,lcsh:Medicine ,Expression ,Biology ,medicine.disease_cause ,Molecular biology ,Rare codons ,law.invention ,03 medical and health sciences ,Linker ,030104 developmental biology ,Infectious Diseases ,Gene cassette ,Escherichia coli BL21 ,law ,medicine ,Recombinant DNA ,Escherichia coli - Abstract
Objective: To demonstrate the sensitivity of expression of fusion genes to existence of a large number of rare codons in recombinant gene sequenced. Methods: Primers for amplification of cholera toxin B, Shiga toxin B and gfp genes were designed by Primer3 software and synthesized. All of these 3 genes were cloned. Then the genes were fused together by restriction sites and enzymatic method. Two linkers were used as a flexible bridge in connection of these genes. Results: Cloning and fusion of cholera toxin B, Shiga toxin B and gfp genes were done correctly. After that, expression of the recombinant gene construction was surveyed. Conclusions: According to what was seen, because of the accumulation of 12 rare codons of Shiga toxin B and 19 rare codons of cholera toxin B in this gene cassette, the expression of the recombinant gene cassette, in Escherichia coli BL21, failed.
- Published
- 2017