摘要
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 inte-grated into the genome of gametophytes of L. japonica, and the expression product showed the ex-pected bioactivity, implying the proper post-transcript modification in haploid gametophyte.
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 inte-grated into the genome of gametophytes of L. japonica, and the expression product showed the ex-pected bioactivity, implying the proper post-transcript modification in haploid gametophyte.
作者
ZHANG YiChen1,2, JIANG Peng1, GAO JiangTao1,3, LIAO JianMin4, SUN ShiJing4, SHEN ZiLong4 & QIN Song1, 1 Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China
2 College of Chemistry and Life Science, Tianjin Normal University, Tianjin 300387, China
3 Graduate University of Chinese Academy of Sciences, Beijing 100049, China
4 Biotechnology Center, China Pharmaceutical University, Nanjing 210009, China
基金
Supported by the National High Technology Research and Development Program of China (Grant Nos. 20060110A4013 and 2006AA100311)
National Natural Science Foundation of China (Grant No. 40506030)
Innovative Key Project of the Chinese Academy of Sciences (Grant No. KZCX2-YW-209)
