Function and application study on prokaryotic enhancer-like element VV1 from vaccinia virus genome
10.3760/cma.j.issn.1003-9279.2015.01.017
- VernacularTitle:痘苗病毒VV1原核增强子样序列的克隆及其功能和应用研究
- Author:
Feng HAN
1
;
Ru CAO
;
Yan WANG
;
Xiaolin BI
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Vaccinia virus;
Sequence analysis
- From:
Chinese Journal of Experimental and Clinical Virology
2015;29(1):50-52
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone enhancer-like sequences from vaccinia virus genome.Study on function and application of it.Methods Enhancer-like element from vaccinia virus genome was obtained by using the chloramphenicol acetyl-transferase(CAT)gene as reporter gene.Stepwise deletion experiment was used to identify the functional domain of VV1 element.Interferon was expressed by using an expression vector harboring VV1 sequence.Results An enhancer-like element VV1 of 283 bp from vaccinia virus genome DNA was obtained.Deletion of β-galactosidase activity showed that positive direction could increase the activity 10.9 times and negative direction could increase 3.8 times.Stepwise deletion experiment was used to identify the functional domain of VV1 element.The results suggested that the 20 bp at 5' terminal and 20 bp at 3' terminal were important to the activity of VV1 and its activity decreased greatly without them.Furthermore,the 30-50 bp at 5' terminal was essentiM to its activity and would lead to complete loss of its activity without it.The antiviral activity of interferon α-2b was increased by 2.6 times in comparison with the original expression plasmid.Conclusion VV1 enhancer-like sequences was obtained from vaccinia virus genome.Stepwise deletion experiment was identification of the functional domain of VV1 element.Interferon gene was highly expressed by using an expression vector harboring enhancer-like sequencesVV1.
