Volume 51, Number 12, 1116-1120, DOI: 10.1007/s11427-008-0143-4

Recombinant expression of rt-PA gene (encoding Reteplase) in gametophytes of the seaweed Laminaria japonica (Laminariales, Phaeophyta)

YiChen Zhang, Peng Jiang, JiangTao Gao, JianMin Liao, ShiJing Sun, ZiLong Shen and Song Qin

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Abstract

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.

Keywords   Laminaria japonica  - gametophyte - genetic engineering - Reteplase - rt-PA - bioreactor

Contributed equally to this work
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), and Innovative Key Project of the Chinese Academy of Sciences (Grant No. KZCX2-YW-209)

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