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作者首先构建了poxA基因突变的鼠伤寒沙门菌。将pMEG-273的BstⅠ-XhoⅠ之间编码Pox A最后270个氨基酸的1018bp片段去除,构建质粒pMEG-279。pMEG-279的1kb XhoⅠ-XbaⅠ片段带有突变的ΔpoxA270等位基因和旁侧序列。将pMEG-279克隆到有R6K复制子的pKNG101的SalⅠ-XbaⅠ位点,得到自杀质粒pMEG-280。
The authors first constructed poxA mutant Salmonella typhimurium. The 1018 bp fragment encoding the last 270 amino acids of Pox A between BstI-XhoI of pMEG-273 was removed to construct plasmid pMEG-279. The 1 kb XhoI-XbaI fragment of pMEG-279 carries the mutated ΔpoxA270 allele and flanking sequences. PMEG-279 was cloned into the SalI-XbaI site of pKNG101 with the R6K replicon to give the suicide plasmid pMEG-280.