Margolin W, Howe M M
Nucleic Acids Res. 1986 Jun 25;14(12):4881-97. doi: 10.1093/nar/14.12.4881.
The C gene of bacteriophage Mu, required for transcription of the phage late genes, was localized by construction and analysis of a series of deleted derivatives of pKN50, a plasmid containing a 9.4 kb Mu DNA fragment which complements Mu C amber mutant phages for growth. One such deleted derivative, pWM10, containing only 0.5 kb of Mu DNA, complements C amber phages and transactivates the mom gene, one of the Mu late genes dependent on C for activation. The DNA sequence of the 0.5 kb fragment predicts a single long open reading frame coding for a 140 amino acid protein. Sequence analysis of DNA containing a C amber mutation located the base change to the second codon of this reading frame. Generation of a frameshift mutation by filling in a BglII site spanning codon 114 of this reading frame resulted in the loss of C complementation and transactivation activity. These results indicate that this open reading frame encodes the Mu C gene product. Comparison of the predicted amino acid sequence of the C protein with those of other transcriptional regulatory proteins revealed some similarity to a region highly conserved among bacterial sigma factors.
噬菌体Mu晚期基因转录所需的C基因,通过构建和分析pKN50的一系列缺失衍生物进行定位,pKN50是一种质粒,含有一个9.4 kb的Mu DNA片段,可补充Mu C琥珀突变噬菌体的生长。一种这样的缺失衍生物pWM10,仅含有0.5 kb的Mu DNA,可补充C琥珀噬菌体并反式激活mom基因,mom基因是依赖C激活的Mu晚期基因之一。0.5 kb片段的DNA序列预测有一个单一的长开放阅读框,编码一个140个氨基酸的蛋白质。对含有C琥珀突变的DNA进行序列分析,将碱基变化定位到该阅读框的第二个密码子。通过填充跨越该阅读框第114密码子的BglII位点产生移码突变,导致C互补和反式激活活性丧失。这些结果表明,这个开放阅读框编码Mu C基因产物。将C蛋白的预测氨基酸序列与其他转录调节蛋白的序列进行比较,发现与细菌σ因子中高度保守的区域有一些相似性。