Zhang YiChen, Jiang Peng, Gao JiangTao, Liao JianMin, Sun ShiJing, Shen ZiLong, Qin Song
Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China.
Sci China C Life Sci. 2008 Dec;51(12):1116-20. doi: 10.1007/s11427-008-0143-4. Epub 2008 Dec 18.
The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte. The expression of foreign genes in sporophte has been proved. In this research, the recombinant expression in gametophyte was investigated by particle bombardment with the rt-PA gene encoding the recombinant human tissue-type plasminogen activator (Reteplase), which is a thrombolytic agent for acute myocardial infarction (AMI). Transgenic gametophytes were selected by their resistance to herbicide phosphiothricin (PPT), and proliferated in an established bubble column photo-bioreactor. According to the results from quantitative ELISA, Southern blotting, and fibrin agarose plate assay (FAPA) for bioactivity, it was showed that the rt-PA gene had been integrated into the genome of gametophytes of L. japonica, and the expression product showed the expected bioactivity, implying the proper post-transcript modification in haploid gametophyte.
海带的生命周期涉及二倍体孢子体和单倍体配子体之间的世代交替。已证实外源基因在孢子体中表达。在本研究中，通过用编码重组人组织型纤溶酶原激活剂（瑞替普酶）的rt-PA基因进行粒子轰击，研究了配子体中的重组表达，瑞替普酶是一种用于急性心肌梗死（AMI）的溶栓剂。通过对除草剂草丁膦（PPT）的抗性筛选转基因配子体，并在已建立的鼓泡柱光生物反应器中增殖。根据定量ELISA、Southern印迹和生物活性纤维蛋白琼脂糖平板试验（FAPA）的结果，表明rt-PA基因已整合到海带配子体基因组中，表达产物显示出预期的生物活性，这意味着在单倍体配子体中进行了适当的转录后修饰。
