Recombination and Heterologous Expression of Allophycocyanin Gene in the Chloroplast of Chlamydomonas reinhardtii

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...

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Bibliographic Details
Published in:Acta biochimica et biophysica Sinica 2005-10, Vol.37 (10), p.709-712
Main Authors: SU, Zhong‐Liang, QIAN, Kai‐Xian, TAN, Cong‐Ping, MENG, Chun‐Xiao, QIN, Song
Format: Article
Language:English
Subjects:
allophycocyanin gene
Animals
Blotting, Southern
Blotting, Western
Chlamydomonas reinhardtii
Chlamydomonas reinhardtii - genetics
Chlamydomonas reinhardtii - metabolism
chloroplast transformation
Chloroplasts - genetics
Chloroplasts - metabolism
Cyanobacteria
DNA, Algal - genetics
Genetic Vectors
Phycocyanin - genetics
Transformation, Genetic - genetics
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Summary: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. Edited by 
 Xue‐Ping ZHOU
ISSN:1672-9145
1745-7270
DOI:10.1111/j.1745-7270.2005.00092.x