Thierbach G, Kalinowski J, Bachmann B, Pühler A
Degussa AG/ASTA Pharma AG, Bielefeld, Federal Republic of Germany.
Appl Microbiol Biotechnol. 1990 Jan;32(4):443-8. doi: 10.1007/BF00903780.
The Corynebacterium glutamicum/Escherichia coli shuttle vector plasmid pZ1 was used to clone the S-(2-aminoethyl)-D,L-cysteine (AEC)-resistance gene from a lysine-excreting, AEC-resistant strain of C. glutamicum, the aspartokinase activity of which was released from feedback inhibition by mixtures of lysine and threonine or AEC and threonine respectively. A recombinant plasmid designated pCS2 carrying a 9.9-kb chromosomal insert that conferred AEC resistance and the ability to excrete lysine to its host was isolated. The aspartokinase activity of the pCS2-carrying strain was resistant towards inhibition by mixtures of lysine and threonine or AEC and threonine respectively. By deletion analysis the DNA region conferring AEC resistance to the host and feedback resistance to its aspartokinase activity could be confined to a 1.2-kb DNA fragment.
利用谷氨酸棒杆菌/大肠杆菌穿梭载体质粒pZ1,从一株分泌赖氨酸且对S-(2-氨基乙基)-D,L-半胱氨酸(AEC)具有抗性的谷氨酸棒杆菌菌株中克隆AEC抗性基因,该菌株的天冬氨酸激酶活性分别被赖氨酸和苏氨酸或AEC和苏氨酸的混合物解除反馈抑制。分离得到一种重组质粒,命名为pCS2,它携带一个9.9kb的染色体插入片段,赋予宿主AEC抗性和分泌赖氨酸的能力。携带pCS2的菌株的天冬氨酸激酶活性分别对赖氨酸和苏氨酸或AEC和苏氨酸的混合物抑制具有抗性。通过缺失分析,可将赋予宿主AEC抗性及其天冬氨酸激酶活性反馈抗性的DNA区域限定在一个1.2kb的DNA片段内。