单增李斯特标准菌NrdD基因插入突变株的构建
摘要
利用同源重组的原理构建了单增李斯特氏菌NrdD基因插入突变株,从大肠杆菌基因组中分别扩增出NrdD基因上下游同源臂,与pKSV7质粒相连,构建成功重组突变载体pKSV7-D1D2,将其电转入单增李斯特菌后,用氯霉素抗性平板筛选得到NrdD基因插入突变株,并对突变菌株序列测定,证明回复突变株构建成功。该突变株可在严格无氧环境中生存,为进一步研究NrdD基因功能、单增李斯特的致病机制和疫苗载体的研发奠定了基础。
引文
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[2]SUN X,HARDER J,KROOK M,et al.A possible glycine radical in anaerobic ribonucleotide reductase from Escherichia coli:nucleotide sequence of the cloned nrdD gene[J].National academy of sciences,1993,90(2):577-581.
[3]OFER A,KREFT J,LOGAN D T,et al.Implications of the inability of Listeria monocytogenes EGD-e to grow anaerobically due to a deletion in the class III NrdD ribonucleotide reductase for its use as a model laboratory strain.[J].Journal of bacteriology,2011,193(12):2931-2940.
[4]周云,潘蕾,郝春秋,等.单增李斯特菌作为疫苗载体的研究进展[J].中国免疫学杂志,2011(12):1132-1134.