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)