Nieradko J, Szewczyk B
Department of Microbiology, University of Gdańsk, Poland.
Acta Biochim Pol. 1988;35(4):357-66.
A fragment of E. coli bacteriophage T4 genome including the four genes (genes 51, 27, 28, 29) coding for the central plug proteins was cloned into plasmid pMCC17. The genes present on this fragment were expressed in E. coli in the absence of phage infection producing hub proteins, which could be identified on polyacrylamide gels. By applying affinity chromatography protein 29 was purified from extracts of E. coli transformed with this hybrid plasmid. The isolated protein had the ability to complement T4 29 amber mutants. The molecular weight of the purified protein was estimated as 75,000 to 85,000 depending on the composition of SDS-polyacrylamide gel used for the assay.
一段包含编码中心塞蛋白的四个基因（基因51、27、28、29）的大肠杆菌噬菌体T4基因组片段被克隆到质粒pMCC17中。该片段上的基因在没有噬菌体感染的情况下在大肠杆菌中表达，产生枢纽蛋白，这些蛋白可以在聚丙烯酰胺凝胶上被鉴定出来。通过亲和层析，从用这种杂交质粒转化的大肠杆菌提取物中纯化出了蛋白29。分离出的蛋白具有互补T4 29琥珀突变体的能力。根据用于分析的SDS-聚丙烯酰胺凝胶的组成，纯化蛋白的分子量估计为75,000至85,000。
