Screening and application of prokaryotic enhancer-like sequence 3A.
- Author:
Feng HAN
1
;
Jian-Xin HE
;
Xiao-Hui LIAO
;
Yan WANG
;
Shu-Hua WU
Author Information
- Publication Type:Journal Article
- MeSH: Enhancer Elements, Genetic; genetics; Escherichia coli; genetics; Gene Expression; drug effects; genetics; Genes, Reporter; Genetic Vectors; chemistry; Interferons; chemistry; genetics; metabolism; Prokaryotic Cells; Sequence Homology, Nucleic Acid; beta-Galactosidase; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2010;24(3):175-177
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSEnhancer-like element from Escherichia coli strain C600 genome was obtained by using the chloramphenicol 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.
RESULTSAn 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 beta-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 interferon alpha-2b was increased by 3.7 times in comparison with the original expression plasmid.
CONCLUSION3A enhancer-like sequence was screened from Escherichia coli strain C600 genome. Interferon gene was highly expressed by using an expression vector harboring enhancer-like sequences.