Yamaguchi A, Someya Y, Sawai T
Division of Microbial Chemistry, Faculty of Pharmaceutical Sciences, Chiba University, Japan.
FEBS Lett. 1993 Jun 14;324(2):131-5. doi: 10.1016/0014-5793(93)81378-d.
The tetA gene was cut into its N- and C-terminal halves at the central EcoRI site and the two halves were subcloned individually or together under a separate lac promoter/operator. The expression of the C-terminal half was detected with a C-terminal-specific antibody. The amount of the N-terminal half in the cytoplasmic membrane was not affected by the presence of the C-terminal half. In contrast, the amount of the C-terminal half in the membrane was increased in the presence of the N-terminal half, indicating that the N-terminal half helps the stable folding of the C-terminal half in the membrane. Each half individually showed no tetracycline transport activity, however, when both halves were expressed together, the resultant complex showed about 40% of the tetracycline transport activity of the wild-type per number of the C-terminals of TetA protein in the membrane.
tetA基因在中央EcoRI位点被切割成N端和C端两半，这两半分别或一起被亚克隆到一个单独的lac启动子/操纵子下。用C端特异性抗体检测C端半段的表达。细胞质膜中N端半段的量不受C端半段存在的影响。相反，在N端半段存在的情况下，膜中C端半段的量增加，这表明N端半段有助于C端半段在膜中稳定折叠。然而，每一半单独都没有四环素转运活性，当两半一起表达时，产生的复合物显示出膜中TetA蛋白每个C端的四环素转运活性约为野生型的40%。
