| Abstract: | Heterogeneous expression of multiple genes in the nucleus of transgenic plants requires the introduction of an individual gene and the subsequent backcross to reconstitute multi-subunit proteins or metabolic pathways.In order to accomplish the expression of multiple genes in a single transformation event,we inserted both large and small subunits of allophycocyanin gene (apcA and apcB) into Chlamydomonas reinhardtii chloroplast expression vector,resulting in papc-S.The constructed vector was then introduced into the chloroplast of C.reinhardtii by micro-particle bombardment.Polymerase chain reaction and Southern blot analysis revealed that the two genes had integrated into the chloroplast genome.Western blot and enzyme-linked immunosorbent assay showed that the two genes from the prokaryotic cyanobacteria could be correctly expressed in the chloroplasts of C.reinhardtii.The expressed foreign protein in transformants accounted for about 2%-3% of total soluble proteins.These findings pave the way to the reconstitution of multi-subunit proteins or metabolic pathways in transgenic C.reinhardtii chloroplasts in a single transformation event. |
