鸡染色质绝缘子HS4的不同区域对杆状病毒表达egfp基因的影响  

The Effect of the Different Regions of Chicken HS4 Insulator on the egfp Gene Expression Mediated by Baculovirus Vectors

作　　者：胡樱虹[1] 钟江[1] 

机构地区：[1]复旦大学生命科学学院微生物学与微生物工程系,上海200433

出　　处：《复旦学报（自然科学版）》2013年第4期442-446,F0002,共6页Journal of Fudan University：Natural Science

基　　金：国家自然科学基金资助项目(30770079)

摘　　要：染色质绝缘子是一种具有保护目的基因免受周围基因调控影响,保证基因稳定表达的DNA元件.前期的研究表明,当鸡染色质绝缘子HS4被置于目的基因表达框下游时,可以显著提高杆状病毒介导的基因的表达水平.为了进一步认识HS4作用的机制,构建了携带HS4绝缘子不同区域的6种重组病毒,比较了它们对报告基因egfp表达的影响.结果发现,相比对照病毒,插入HS4全长片段的重组病毒vBG-HS4-1200,以及同时插入250bp核心区域和400bp连接片段的重组病毒vBG-HS4-650在感染Sf9细胞后报告基因egfp的表达水平明显提高.这个结果与前人在哺乳细胞表达系统中所得出的结果基本相符,从而进一步证实HS4在裂解型病毒中同样可以通过表观遗传学修饰调控基因的表达.Chromatin insulators are DNA elements that are able to maintain the stable expression of target gene by isolating it from regulation of surrounding regulatory factors. Previous research showed that chicken ehromatin insulator HS4 significantly improved baculovirus-mediated gene expression when it was placed downstream of the gene expression cassette. In order to identify the part of HS4 that is essential to improve baculovirus-mediated gene expression, six recombinant baeulovirus vectors containing different regions of HS4 insulator downstream the expression cassette of reporter gene, egfp, were constructed. The expression of EGFP was significantly improved in cells infected by vBG-HS4 650 and vBG-HS4-1200, which contained the combined the 250 bp 5＇core and the 400 bp 3＇distal region of HS4, and the full-length HS4, respectively, compared with the control virus vBG. This result is consistent with findings in mammalian cells, and showed that epigenetie regulation might play an important role in viral gene expression during baeulovirus lyric infection.

关 键 词：杆状病毒 基因表达 绝缘子 

分 类 号：Q812[生物学—生物工程]