Su W C, Tung S Y, Yang M K, Kuo T T
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, Republic of China.
Mol Gen Genet. 1999 Aug;262(1):22-6. doi: 10.1007/s004380051055.
Host factors that are important for infection of Xanthomonas campestris pv. citri by the filamentous bacteriophage cf were investigated by transposon mutagenesis with Tn5tac1. A mutant, XT501, that was resistant to cf infection was recovered, showing that the gene inactivated by the transposon is required for infection by the phage but not for cf replication or assembly. A 1.7-kb SacI-ApaI DNA fragment from XT501 containing the bacterial DNA flanking one end of the transposon was cloned and shown to be required for cf infection. Nucleotide sequence analysis of the 1.7-kb fragment reveals the presence of an ORF that encodes a protein of 146 amino acids. This protein shows 42% identity to the type 4 prepilin encoded by the pilA genes of other bacteria. The pilA gene of X. campestris pv. citri is thus essential for infection by the bacteriophage cf.
利用Tn5tac1转座子诱变技术,研究了对丝状噬菌体cf感染野油菜黄单胞菌柑橘致病变种(Xanthomonas campestris pv. citri)至关重要的宿主因子。获得了一株对cf感染具有抗性的突变体XT501,这表明转座子使其失活的基因是噬菌体感染所必需的,但对cf的复制或组装并非必需。从XT501中克隆了一个1.7kb的SacI - ApaI DNA片段,该片段含有位于转座子一端侧翼的细菌DNA,并且已证明它是cf感染所必需的。对该1.7kb片段的核苷酸序列分析揭示了一个编码146个氨基酸蛋白质的开放阅读框(ORF)。该蛋白质与其他细菌pilA基因编码的4型前菌毛蛋白具有42%的同一性。因此,野油菜黄单胞菌柑橘致病变种的pilA基因对于噬菌体cf的感染至关重要。