摘要
目的建立鸡沙门菌NCTC13346株基因组物理图谱,并比较其结构特征。方法制备鸡沙门菌NCTC13346株基因组DNA,用rrl基因特异性的I-CeuⅠ内切酶以及限制性内切酶XbaⅠ和AvrⅡ酶切后,进行脉冲凝胶电泳(PFGE),分离出DNA片段,最后用噬菌体P22转导已知携带沙门氏菌基因的Tn10插入子的方法定位基因排列,根据酶切片断的连接与已知基因的定位,绘制该菌的基因组结构物理图,并与已知鸡沙门菌SARB21基因组结构进行比较。结果建立鸡沙门菌NCTC13346株基因组物理图谱,与鸡沙门菌SARB21基因组物理图谱相比,两者具有相对于鼠伤寒沙门氏菌LT2的相似的基因插入和缺失片段,但由于两者基因组中含有rrn操纵子的DNA片段的不同排列,分别表现为迥异的基因组结构。结论建立鸡沙门菌基因组物理图谱可用于分析同种细菌的基因组特点与进化。
Objective To construct a physical map and compare the character on the genomic structure of S. gallinarum NCTC13346.Methods The genomic DNA isolation from S. gallinarum NCTC13346 were cleavage with Xba Ⅰ , Avr Ⅱor I-Ceu Ⅰ and the fragments of genome were separated using the Pulsed field gel electrophoresis（PEGE）. Then, the bacteriophage P22 intermediated transduction of Tn10 insertion bring the known genes was also employed for locating genes and refining the physical map, and the comparative analysis was carried between the genome of S. gallinarum NCTC13346 and SARB21. Results The genome physical map of S. gallinarum NCTC13346 was constructed, whichshares most insertions and deletions similar with strain SARB21 relative to S. typhimurium LT2,whereas,those differences in the arrangement of genomic DNA segments flanking by the rm operons, what leading to distinct genome structures of the S. gallinarum. Conclusion The construction of a genomic physical map of bacteria is a useful method for analysis of the evolution and genomic character of bacteria in the same species.
出处
《哈尔滨医科大学学报》
CAS
北大核心
2006年第6期450-453,共4页
Journal of Harbin Medical University
