[Screening and application of enhancer-like sequences from vaccinia virus].
OBJECTIVE
To screen enhancer-like sequences from vaccinia virus genome, to construct an expression vector harboring prokaryotic enhancer-like sequence and study the effect of interferon gene expression.


METHODS
Enhancer-like element from vaccinia virus genome was obtained by using the chloramphenicol acetyl-transferase cat gene as reporter gene. An expression vector harboring prokaryotic enhancer-like sequence VV1 from vaccinia virus was constructed. Interferon was expressed and assayed.


RESULTS
Eighteen enhancing sequences were found. From them two enhancer-like sequences with distance and orientation independence property were screened and named VV1 and VV16 respectively. Quantification test showed that the direct and reverse orientation of VV1 could increase the activity of beta-galactosidase with 10.9 and 3.8 times and those of VV16 could increase by 9.0 times and 4.1 times respectively. The enhancing activity of the element was on transcription level. An expression vector harboring prokaryotic enhancer-like sequence VV1 was constructed. Using this vector the antiviral activity of interferon alpha-2b was increased by 2.6 times in comparison with the original expression plasmid.


CONCLUSIONS
Two enhancer-like sequences were screened from vaccinia virus genome. Interferon gene was highly expressed by using an expression vector harboring enhancer-like sequences.
Han
Feng
F
Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Bi
Xiao-lin
XL
Cao
Ru
R
Wang
Yan
Y
Wu
Shu-hua
SH
D004742:Enhancer Elements, Genetic; D005822:Genetic Vectors / Q000235:genetics*; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D010957:Plasmids; D011994:Recombinant Proteins; D014616:Vaccinia virus / Q000235:genetics*
D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins