CAS and Shandong Province Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China.
Mar Drugs. 2021 May 28;19(6):315. doi: 10.3390/md19060315.
, a unicellular marine green alga, is used widely in aquaculture as an initial feeding for fish, bivalve mollusks, penaeid shrimp larvae, and rotifers because of its rich content of amino acids and fatty acids. A stable nuclear transformation system using the herbicide phosphinothricin (PPT) as a selective reagent was established previously. In this research, the recombinant expression in was investigated by particle bombardment with the gene that encodes the recombinant human tissue-type plasminogen activator (Reteplase), which is a thrombolytic agent for acute myocardial infarction treatment. Transgenic algal strains were selected by their resistance to PPT, and expression of was validated by PCR, Southern blotting, and Western blotting, and bioactivity of rt-PA was confirmed by the fibrin agarose plate assay for bioactivity. The results showed that was integrated into the genome of , and the expression product was bioactive, indicating proper post-transcriptional modification of rt-PA in . This report contributes to efforts that take advantage of marine microalgae as cell factories to prepare recombinant drugs and in establishing a characteristic pathway of oral administration in aquaculture.
, 一种单细胞海洋绿藻，由于其富含氨基酸和脂肪酸，被广泛应用于水产养殖中，作为鱼类、双壳贝类、对虾幼体和轮虫的初始饲料。先前已经建立了一种使用除草剂草丁膦（PPT）作为选择试剂的稳定核转化系统。在这项研究中，通过粒子轰击法用编码重组人组织型纤溶酶原激活剂（Reteplase）的 基因进行了 的重组表达，Reteplase 是一种用于治疗急性心肌梗死的溶栓剂。通过对 PPT 的抗性选择转基因藻类品系，并通过 PCR、Southern 印迹和 Western 印迹验证 表达，通过纤维蛋白琼脂糖平板测定生物活性确认 rt-PA 的生物活性。结果表明， 整合到 的基因组中，表达产物具有生物活性，表明 rt-PA 在 中得到了适当的转录后修饰。本报告有助于利用海洋微藻作为细胞工厂来制备重组药物，并在水产养殖中建立口服给药的特征途径。
