将氯霉素(Cm)抗性基因克隆到pUTmini-Tn5Km2载体中,利用含卡那霉素(Km)和氯霉素(Cm)抗性基因的pUTmini-Tn5Km2(Cm)转座载体将Km和Cm抗性基因随机插入鸡白痢沙门氏菌基因组中,以相应的抗生素进行筛选,获得大量在不同位点插入突变的突变体,从中筛选鸡白痢沙门氏菌某功能缺陷型突变株。通过对突变株的基本特征以及PCR鉴定后,再进行插入基因定位。结果表明,Km和Cm抗性基因已成功转座至鸡白痢沙门氏菌基因组上;基因定位显示突变株均有且只有1个插入位点,插入位点的位置不尽相同。这为研究鸡白痢沙门氏菌功能基因和筛选特定突变株提供了必要的基础。
Cm^R and Km^R genes were transposed into genomic DNA of Salmonella pullorum to produce a library of mutants of Salmonella pullorum by pUTmini-TnSKm2 (Cm) which was constructed by inserting CmR gene into pUTmini-TnSKm2. Then some defective mutants were screened. Mutants were identified by basic characteristic of Salmonella pullorum, special PCR and gene location. The results showed that CmR and KmR genes were integrated into genome of Salmonella pullorum mutants and each mutant has one and only one different locus of transposon in the genome. These results provided essential base for studying the function gene of Salmonella pullorum and screening special mutants.