Pankova S V, Abilev S K
Genetika. 1993 Mar;29(3):539-42.
Cloning the genes for threonine biosynthesis from Propionibacterium shermanii was performed in Escherichia coli cells using the plasmid vector pVZ361. The cloned genes were identified via complementation of thrB, thrC, thrA1 and thrA2 mutations of E. coli. The gene complementing thrB of E. coli was located within a 5.1 kb fragment of P. shermanii chromosomal DNA. The cloned DNA fragment hybridized with a chromosomal fragment of P. shermanii of the same size. The plasmid pSPt4 (with the thrB gene) was digested with Sau3A and ligated with the BamHI-restricted pUC19 vector. The 1.8 kb DNA fragment of P. shermanii was shown to complement the thrB gene function in E. coli cells.
利用质粒载体pVZ361在大肠杆菌细胞中克隆了谢氏丙酸杆菌苏氨酸生物合成基因。通过对大肠杆菌thrB、thrC、thrA1和thrA2突变的互补作用鉴定出克隆的基因。互补大肠杆菌thrB的基因位于谢氏丙酸杆菌染色体DNA的一个5.1 kb片段内。克隆的DNA片段与大小相同的谢氏丙酸杆菌染色体片段杂交。用Sau3A酶切质粒pSPt4(含thrB基因)并与经BamHI酶切的pUC19载体连接。结果表明,谢氏丙酸杆菌的1.8 kb DNA片段可在大肠杆菌细胞中互补thrB基因功能。