Maksimoa T G, Mustaev A A, Zaĭchikov E F, Baranova L V, Kumarev V P
Bioorg Khim. 1989 Jan;15(1):18-23.
A highly selective affinity label was introduced into the T7 phage RNA polymerase by means of GMP ortho-formylphenyl ester and [alpha-32P]UTP nearby the enzyme's active site, which was located using limited cleavage technique. Hydroxylamine, bromine, N-chlorosuccinimide, and cyanogen bromide were employed as the reagents. Analysis of gel-electrophoretic patterns of the cleavage products led to a conclusion that Lys631 is the target of labelling. The region nearby this residue has a high degree of sequence homology with regions of RNA polymerases from T3 and SP6 phages and yeast mitochondria.
通过在T7噬菌体RNA聚合酶活性位点附近引入鸟苷酸邻甲酰苯酯和[α-32P]UTP，构建了一种高选择性亲和标记物，该活性位点通过有限切割技术定位。使用羟胺、溴、N-氯代琥珀酰亚胺和溴化氰作为试剂。对切割产物的凝胶电泳图谱分析得出结论，Lys631是标记的靶点。该残基附近的区域与T3和SP6噬菌体以及酵母线粒体的RNA聚合酶区域具有高度的序列同源性。
