Expression of synthetic calcitonin genes in plasmid vectors containing tandemly repeated non-overlapping ribosome binding sites
- PMID: 2480257
- DOI: 10.1016/0020-711x(89)90231-0
Expression of synthetic calcitonin genes in plasmid vectors containing tandemly repeated non-overlapping ribosome binding sites
Abstract
1. Expression plasmids containing non-overlapping tandemly repeated ribosome binding sites (RBS) were constructed in order to stabilize mRNA and enhance translation. 2. Two synthetic genes (human calcitonin tetramer gene and a fusion gene human gamma-interferon-human calcitonin) were cloned in these vectors and the effect of multiplicity of Shine-Dalgarno (S/D) sequence on heterologous gene expression was studied. 3. It was found that duplication and triplication of RBS had no effect on the stability of mRNA but led to a strong decrease in the level of recombinant protein and mRNA in the cell. 4. Plasmids bearing four times repeated S/D sequences gave longer-lived mRNAs and maintained a level of protein and mRNA very close to the values obtained with a single S/D containing plasmids.
Similar articles
-
Expression of repetitive human calcitonin genes in Escherichia coli.Biotechnol Appl Biochem. 1989 Aug;11(4):401-12. Biotechnol Appl Biochem. 1989. PMID: 2673299
-
Gene expression evidence indicates that nucleotides 507-513 and 1434-1440 in 16S rRNA are organized in close proximity on the Escherichia coli 30S ribosomal subunit.Eur J Biochem. 1997 Aug 15;248(1):10-4. doi: 10.1111/j.1432-1033.1997.00010.x. Eur J Biochem. 1997. PMID: 9310353
-
The use of two-cistron constructions in improving the expression of a heterologous gene in E. coli.Nucleic Acids Res. 1990 Apr 11;18(7):1711-8. doi: 10.1093/nar/18.7.1711. Nucleic Acids Res. 1990. PMID: 2110654 Free PMC article.
-
Vectors with adjustable copy numbers.Biotechnology. 1988;10:227-35. doi: 10.1016/b978-0-409-90042-2.50017-9. Biotechnology. 1988. PMID: 3061515 Review. No abstract available.
-
High-expression of a target gene and high-stability of the plasmid.Appl Biochem Biotechnol. 1991 Feb;27(2):145-62. doi: 10.1007/BF02921523. Appl Biochem Biotechnol. 1991. PMID: 2029184 Review. No abstract available.
Cited by
-
Effects of a recombinant gene expression on ColE1-like plasmid segregation in Escherichia coli.BMC Biotechnol. 2011 Mar 1;11:18. doi: 10.1186/1472-6750-11-18. BMC Biotechnol. 2011. PMID: 21362179 Free PMC article.
-
Increasing signal specificity of the TOL network of Pseudomonas putida mt-2 by rewiring the connectivity of the master regulator XylR.PLoS Genet. 2012;8(10):e1002963. doi: 10.1371/journal.pgen.1002963. Epub 2012 Oct 11. PLoS Genet. 2012. PMID: 23071444 Free PMC article.