DOI: 10.3724/SP.J.1249.2016.01033
Lpaat from Brassica napus and gpd1 from Saccharomyces cerevisiae were optimized according to the codon of Chlamydomonas reinhardtii to obtain c-lpaat and c-gpd1. The modified DNA fragments were amplified and inserted into plasmid pH124 to construct Chlamydomonas expression vectors. The expression vectors were then introduced into Chlamydomoas with the glass-bead-method and the transgenic algae containing c-lpaat and c-gpd1 was selected after screening with zeomycin resistance and polymerase chain reaction(PCR)analysis. Finally, the transgenic algae in which c-lpaat and c-gpd1 were expressed effectively was screened by semi-quantitative reverse transcription-PCR(RT-PCR). The transcription level of c-lpaat and c-gpd1 in transgenic algae was increased by 56.38% and 75.95%, respectively, resulting in correspondingly increased lipid content. In conclusion, the exogenous genes from the plants could be effectively expressed in Chlamydomonas reinhardtii and the transgenic algae with high lipid could be screened quickly by semi-quantitative RT-PCR. These results lay the foundation of quickly screening for transgenic algae with high lipid content.
