偶氮苯修饰的DNA对引物延伸的光调控

Photoregulating primer extension using azobenzene linked DNA template

研究偶氮苯单元修饰在核酸上控制引物延伸的行为。系统地筛选5、6、7和8个碱基的保护链通过4,4′-二羟甲基偶氮苯连接的25 mer DNA模板,并研究其在紫外光照前后调控的引物延伸效率。结果表明,具有7个保护碱基的C3对Pri.15和6个碱基短链的C2对Pri.17都具有较好的光调控延伸效果。其中C3,在Vent酶作用下紫外光照后引物延伸效率增加1倍以上。而C2,尽管紫外光照前增加了引物延伸的背景,在Vent酶催化下紫外光照射后的延伸产率达到91.4%,比紫外光照前增加84%。本工作为分子水平上研究基因功能、基因表达网络以及疾病的发生和发展提供了一种新的策略和研究手段。

It has been a hot topic in recent years to carry out a series of life activities by artificial reversible manipulation of specific gene expression. In this work, the photocontrol of primer extension directed by azobenzene modified DNA templates was investigated. DNA templates attached by protective ODNs with 5, 6, 7, and 8 complementary bases through 4,4′-bis(hydroxymethyl)azobenzene were systematically evaluated in photoregulation of the primer extension without or with UV irradiation. The results showed high efficiencies of C3 with 7 protected bases and C2 with 6-base short chains for photoregulating Pri.15 and Pri.17, respectively. Especially, for C3, when catalyzed by Vent DNA polymerase, primer extension efficiency had one-fold increase upon UV irradiation. Similarly, for C2, the extension efficiency reached 91.4% with UV irradiation and achived 84% increase, compared to that without UV irradiation, although the background of primer extension in the dark was a little high. This work provides a new strategy or means for studying the gene function and gene expression network and for exploring the occurrence and development of disease at the molecular level.