Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry of Vilnius University, Mokslininkų 12, 08662, Vilnius, Lithuania.
Arch Virol. 2014 Feb;159(2):327-31. doi: 10.1007/s00705-013-1800-x. Epub 2013 Aug 15.
The bacteriophage T4 insertion-substitution (I/S) vector system has become one of the most important tools for the introduction of site-directed mutations into the T4 genome. In this study, we show that the I/S phage T4 K10 carries two point mutations within the gene for polynucleotide kinase pseT, resulting in amino acid substitutions G14D and R229H. The G14D mutation impairs 5'-kinase activity in vivo as well as in vitro and leads to diminished processing at secondary sites of several RegB-cleaved transcripts.
T4 噬菌体插入-取代（I/S）载体系统已成为向 T4 基因组中引入定点突变的最重要工具之一。在本研究中，我们表明 I/S 噬菌体 T4 K10 在多核苷酸激酶 pseT 基因内携带两个点突变，导致氨基酸取代 G14D 和 R229H。G14D 突变会损害体内和体外的 5'-激酶活性，并导致几种 RegB 切割转录物的次要位点的加工减少。
