Kozlov Iu I, Kochetova L P, Livshits V A, Mashko S V, Moshentseva V N
Genetika. 1980;16(1):66-77.
A set of hybrid plasmids carrying Escherichia coli threonine genes was obtained and cloned. The plasmid pBR322 was used as a vehicle. The genetic and restriction analyses showed that genes thrA and thrB were placed between SalGI and EcoRI sites on the 2.6 megadaltons DNA region. The transcription of threonine operon genes inserted in the hybrid plasmids is under the control of its own promoter. The copy number of hybrid plasmids was reverse proportional to their molecular weight and did not depend on the replicon number. Amplification of genes of threonine operon by hybrid plasmids led to 20-25-fold increase of homoserine dehydrogenase activity, encoded by thrA gene. The expression of this gene, incorporated in hybrid plasmids, was repressed by the addition of threonine and isoleucine in the culture medium.
获得并克隆了一组携带大肠杆菌苏氨酸基因的杂种质粒。使用质粒pBR322作为载体。遗传和限制酶切分析表明,thrA和thrB基因位于2.6兆道尔顿DNA区域的SalGI和EcoRI位点之间。插入杂种质粒中的苏氨酸操纵子基因的转录受其自身启动子的控制。杂种质粒的拷贝数与其分子量成反比,且不依赖于复制子数。杂种质粒对苏氨酸操纵子基因的扩增导致由thrA基因编码的高丝氨酸脱氢酶活性增加了20至25倍。培养基中添加苏氨酸和异亮氨酸可抑制杂种质粒中该基因的表达。