Schnell Robert, Abdulkarim Farhad, Kálmán Miklós, Isaksson Leif A
Department of Microbiology, Stockholm University, S-106 91 Stockholm, Sweden.
FEBS Lett. 2003 Mar 13;538(1-3):139-44. doi: 10.1016/s0014-5793(03)00159-5.
An Escherichia coli strain was constructed in which both chromosomal genes encoding elongation factor (EF)-Tu (tufA and tufB) have been inactivated with precise coding sequence replacements. A tufA gene in an expression vector is supplied as the sole EF-Tu source. By using plasmid replacement, based on plasmid incompatibility, mutant EF-Tu variants with a large C'-terminal extension up to 270 amino acids were studied and proved to be functional in a strain lacking the chromosomal tufA and tufB genes.
构建了一种大肠杆菌菌株,其中编码延伸因子(EF)-Tu的两个染色体基因(tufA和tufB)已通过精确的编码序列替换而失活。表达载体中的tufA基因作为唯一的EF-Tu来源提供。通过基于质粒不相容性的质粒替换,研究了具有长达270个氨基酸的大C末端延伸的突变EF-Tu变体,并证明其在缺乏染色体tufA和tufB基因的菌株中具有功能。