Effects of an anti-alpha monoclonal antibody on interaction of Escherichia coli RNA polymerase with lac promoters.
Effects of an anti-alpha monoclonal antibody on interaction of Escherichia coli RNA polymerase with lac promoters.
复制标题
抗 α 单克隆抗体对大肠杆菌 RNA 聚合酶与 lac 启动子相互作用的影响。
DOI：
10.1021/bi00470a026
复制
发表时间：
1990
期刊：
影响因子：
2.9
通讯作者：
Krakow,JS
中科院分区：
文献类型：
--
作者：
Riftina,F;DeFalco,E;Krakow,JS
材料与方法试剂如下获得：cAMP，阿帕，1 Sigma Chemical Co.; d（AT）n，核糖核苷三磷酸，Pharmacia;[3 H] UTP，[y-32 P] ATP，和[a-32 P] dATP，ICN; T4多核苷酸激酶，碱性磷酸酶，DNA聚合酶I Klenow片段，和限制性内切核酸酶，BoehringerMannheim; DNA酶I，库珀生化;尿素，Schwarz-Mann;硫酸二甲酯，哌啶，和肼，Aldrich;甲酰胺，Amresco;丙烯酰胺，Serva; Snintisol，Isolab. RNA聚合酶和抗体的制备。从大肠杆菌中纯化RNA聚合酶。coliK 12细胞，通过修改Burgess和Jendrisak（1975）的方法。如Lowe等人所述，通过变性小牛胸腺DNA-琼脂糖层析分离全酶。（1979年）。RNA聚合酶全酶浓度测定
Materials and MethodsMaterials. Reagents were obtained as follows: cAMP, ApA, 1 Sigma Chemical Co.; d (AT)„, ribonucleoside tri-phosphates, Pharmacia;[3H] UTP,[y-32P] ATP, and [a-32P] dATP, ICN; T4 polynucleotide kinase, alkaline phosphatase, DNA polymerase I Klenow fragment, and restriction endonucleases, BoehringerMannheim; DNase I, Cooper Biochemical; urea, Schwarz-Mann; Dimethyl sulfate, piperidine, and hydrazine, Aldrich; formamide, Amresco; acryl-amide, Serva; Scintisol, Isolab.Preparation of RNA Polymerase and Antibody. RNA polymerase was purified from E. coli K12 cells by a modification of the procedure of Burgess and Jendrisak (1975). Holoenzyme was isolated by chromatography on denatured calf thymus DNA-agarose as described by Lowe et al.(1979). RNA polymerase holoenzyme concentration was determined