痘苗病毒增强子样序列的筛选及应用研究  

Screening and application of enhancer-like sequences from vaccinia virus

作　　者：韩峰[1] 秘晓林[1] 曹茹[1] 王艳[1] 吴淑华[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京100052

出　　处：《中华实验和临床病毒学杂志》2007年第4期301-303,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家科委"863"高科技资助项目(102-11-03-03)

摘　　要：目的从痘苗病毒基因组中筛选原核增强子样序列,构建携带原核增强子样序列的表达载体,探讨其对干扰素基因表达的影响。方法采用氯霉素乙酰转移酶基因(cat)作为报告基因,从痘苗病毒基因组中筛选具有原核增强子样活性的序列,构建携带增强子样序列VV1的表达载体,表达干扰素基因和检测干扰素活性。结果从痘苗病毒天坛株DNA基因组中筛选出18个在大肠埃希菌中具有增强活性的序列,从中筛选到两个原核增强子样序列VV1和VV16,VV1正反向分别可使lacZ基因活性提高10.9倍和3.8倍,VV16正反向分别可使lacZ基因活性提高9.0倍和4.1倍;证实它们的增强活性体现在转录水平;用痘苗病毒增强子样序列VV1构建的表达载体,其表达的IFN-α2b型干扰素比原表达载体活性高2.6倍。结论从痘苗病毒天坛株DNA基因组中筛选到2个原核增强子样序列-携带有痘苗病毒增强子样序列的表达载体可提高干扰素基因的表达水平。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 13-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 a-2b was increased by 2.6 times in comparison with the original expression plasmid. Conclusion Two enhancer-like sequences were screened from vaccinia virus genome. Interferon gene was highly expressed by using an expression vector harboring enhancer-like sequences.

关 键 词：痘苗病毒 原核增强子样序列 表达载体 

分 类 号：R346[医药卫生—基础医学]