A New Tandem Gene Construction Method Involving a Cloning System Using Poxvirus DNA polymerase, and Its Application to Gene ExpressionReport as inadecuate




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GFP expressiongene in Escherichia coli weresequentially constructed. Efficacies of the GFP gene expression of those plasmids in E.coli showed an increasing trend in accordance with the copy numbers of thegene. On SDS polyacrylamide gel electrophoresis with Coomassie blue staining,no expressed protein could be seen in E.coli cells harboring plasmids that contained one or two copies of the gene.However, expressed protein bands in E.coli cells were clearly detected with 4 copies of the gene. In quantitativeanalyses involving green fluorescence intensities per culture volume, the expressionlevel in E. coli with 16 copies ofthe gene was 36.3-fold higher than that in E.coli with one copy at 22 hours after induction.

KEYWORDS

Poxvirus DNA polymerase, Gene Cloning, Tandem Repeat, GFP, T7 RNA Polymerase

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Shibui, T. , Sakaguchi, D. and Hara, H. 2014 A New Tandem Gene Construction Method Involving a Cloning System Using Poxvirus DNA polymerase, and Its Application to Gene Expression. Advances in Bioscience and Biotechnology, 5, 838-845. doi: 10.4236-abb.2014.510098.





Author: Tatsuro Shibui*, Daisuke Sakaguchi, Hiroyoshi Hara

Source: http://www.scirp.org/



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