环辛炔修饰的脱氧尿苷三磷酸在DNA中的酶促掺入及荧光标记
Enzymatic incorporation and fluorescent labelling of cyclooctyne-modified deoxyuridine triphosphates in DNA.
作者信息
Ren Xiaomei, Gerowska Marta, El-Sagheer Afaf H, Brown Tom
机构信息
Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Oxford OX1 3TA, UK.
School of Chemistry, University of Southampton, Highfield, Southampton SO17 1BJ, UK.
出版信息
Bioorg Med Chem. 2014 Aug 15;22(16):4384-90. doi: 10.1016/j.bmc.2014.05.050. Epub 2014 Jun 5.
The amino group of 5-aminopropargyl-2'-deoxyuridine-5'-triphosphate was labelled with dibenzocyclooctyne (DIBO) and two derivatives of bicyclo [6.1.0] non-4-yne (BCN) with short and long linkers to produce three different cycloalkyne-modified deoxyuridine triphosphates. BCN was successfully incorporated into DNA at multiple sites by enzyme-mediated primer extension and the polymerase chain reaction (PCR). Efficient fluorescent labelling of the BCN-DNA and DIBO-DNA with Cy3-azide was demonstrated.
5-氨基丙炔基-2'-脱氧尿苷-5'-三磷酸的氨基用二苯并环辛炔（DIBO）以及带有短连接子和长连接子的双环[6.1.0]壬-4-炔（BCN）的两种衍生物进行标记，以产生三种不同的环炔修饰的脱氧尿苷三磷酸。通过酶介导的引物延伸和聚合酶链反应（PCR），BCN成功地在多个位点掺入到DNA中。结果表明，用Cy3-叠氮化物对BCN-DNA和DIBO-DNA进行了高效荧光标记。
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