Screening and application of prokaryotic enhancer-like sequence 3A

Objective To screen enhancer-like sequences from Escherichia coli strain c600 genome,to construct an expression vector harboring prokaryotic enhancer-like sequence and study the effect of interferon gene expression.Methods Enhancer-like element from Escherichia coli strain C600 genome was obtained by using the ehloramphenieol acetyl-transferase(CAT)gene as reporter gene.An expression vector harboring prokaryotic enhancer-like sequence from Escherichia coli strain C600 was constructed.Interferon was expressed and assayed.Results An enhancer-like sequences with distance and orientation independence property were screened and named 3A.Quantification test showed that the direct and reverse orientation of 3A could increase the activity of β-galactosidase with 7.11 and 2.93 times.The enhancing activity of the element was on transcription level.An expression vector harboring the prokaryotic enhancer-like sequence 3P3 which was enhancing function region of sequence 3A was constructed. Using this vector the antiviral activity of interferona-2b was increased by 3.7 times in comparison with the original expression plasmid.Conclusion 3A enhancer-like sequence was screened from Escherwhia coli strain C600 genome.Interferon gene was highly expressed by using an expression vector harboring enhancer-like sequences. Key words: Porkaryotic enbancer-like sequence: Genetic vectors