Construction of Rice Site-Specific Chloroplast Transformation Vector and Transient Expression of EGFP Gene in Dunaliella Salina
Chloroplast is a new hotspot in the field of plant transformation system of plant genetic engineering. Plastid transformation has several advantages: high expression, multiple expressed genes in a single transformation event, absence of gene silencing, et al. A series of elements for
construction of dicistronic site-specific integration expression vector of rice chloroplast have been cloned, including trnI-trnA (rice chloroplast homologous recombination fragments), Prrn (16S rRNA operon promotor), PpsbA (the 3′ untranslated region of the chloroplastpsbA
gene), hptII gene (encoding hygromycin phosphotransferase) and EGFP (encoding enhanced green fluorescence protein). All the elements were constructed into a rice chloroplast dicistronic expression vector pCTE04 (-trnI-Prrn-RBS-hptII-RBS-EGFP-PpsbA- trnA-).
Then pCTE04 was introduced into chloroplasts of Dunaliella salina through particle bombardment. Strong green fluorescence was observed in chloroplasts of some bombarded Dunaliella salina cells under a stereo fluorescence microscope, indicating that pCTE04 could be expressed in
Dunaliella salina chloroplasts transiently. It provides a solid foundation for further genetic engineering in rice chloroplast transformation.
Keywords: CHLOROPLAST; DUNALIELLA SALINA; EGFP; TRANSIENT EXPRESSION
Document Type: Research Article
Publication date: 01 December 2011
- Journal of Biomedical Nanotechnology (JBN) is a peer-reviewed multidisciplinary journal providing broad coverage in all research areas focused on the applications of nanotechnology in medicine, drug delivery systems, infectious disease, biomedical sciences, biotechnology, and all other related fields of life sciences.
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