College of Life Sciences, Yantai University, Yantai 264000, China.
Key Laboratory of Coastal Biology and Biological Resource Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, China.
Mar Drugs. 2022 Jul 28;20(8):484. doi: 10.3390/md20080484.
To solve the problem of antibiotic abuse in aquaculture and to utilize the application potential of antimicrobial peptides (AMPs), a chloroplast transformation system of was successfully constructed for effectively expressing two exogenous AMPs. The endogenous fragments of 16S rDNA/-23S rDNA were used as flanking fragments for the homologous recombination in the chloroplast genome. Two AMPs encoded by the transformation vector were controlled by the native promoter in a polycistron. The plasmids were transferred into via particle bombardment and the transformation vectors were screened using phosphinothricin (), a dominant selection marker under the control of the promoter. Subsequently, in the positive transformed colonies, the exogenous fragments were found to be inserted in the flanking fragments directionally as expected and two foreign AMPs were successfully obtained. Finally, two exogenous peptides with antibacterial properties were obtained from the transformed strain. The two AMPs expressed by the transformed strain were shown to have similar inhibitory effects to antibiotics by inhibition tests. This suggested that AMPs can be introduced into aquaculture using baited microalgae, providing new ideas and ways to solve a series of aquaculture diseases caused by bacteria.
为了解决水产养殖中抗生素滥用的问题，并挖掘抗菌肽（AMPs）的应用潜力，本研究成功构建了 的叶绿体转化体系，以有效表达两种外源性 AMPs。叶绿体基因组中的同源重组使用 16S rDNA/-23S rDNA 的内源性片段作为侧翼片段。由转化载体编码的两种 AMPs 由天然启动子 控制，在多顺反子中表达。质粒通过粒子轰击转入 ，并利用草丁膦（）筛选转化载体，该筛选标记由 启动子控制，为显性选择标记。随后，在阳性转化菌落中，预期的外源片段定向插入侧翼片段中，成功获得了两种外源性 AMPs。最后，从转化株中获得了两种具有抗菌活性的外源性肽。转化株表达的两种 AMPs 通过抑菌试验显示出与抗生素相似的抑制作用。这表明可以利用诱饵微藻将 AMPs 引入水产养殖，为解决一系列由细菌引起的水产养殖疾病提供了新的思路和方法。
