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Abstract To research the quickly construction of universal plant expression vector with different objectives, one expression cassette was constructed with partial sequence of multiple cloning site cloned from pUC19, CaMV 35S promoter and NOS terminator cloned from plasmid pEGAD. The whole expression cassette was inserted into the multiple cloning site of vector pCAMBIA1300 to generate the universal plant expression vector pCamE (GenBank accession No. JX841315). Plasmid pCamE offerred 8 restriction endonuclease sites at the multiple cloning site, 4 upstream of the promoter and 3 downstream of the terminator. These restriction endonuclease sites were helpful to insert foreign gene into vector pCamE, and modify or replace promoter/terminator by any other interested elements. With the report gene of enhanced green fluorescent protein (GFP), 3 expression vectors, pCam::GFP, pCam::SalGFP and pCam::DAHPS-GFP, were constructed and transformed into onion (Allium cepa L.) epidermal cells and Arabidopsis thaliana, respectively. The proteins of eGFP and DAHPS-GFP were expressed normally in onion epidermal cells. In GFP transformed Arabidopsis, eGFP fluorescence was clearly observed in root, root hair, spire base, epidermal hair and anther. The results showed that this universal plant expression vector pCamE could integrate into the plant genome and normally express with different genes, the length of the inserted gene sequence and the site that target gene inserted into did not affect the transcription or expression of these target genes. All these demonstrated that vector pCamE was an economic and wide versatility vector, by which the target gene could be modified efficiently and inherit stablly in transgenic plant with different objectives. Thereby, pCamE provides a powerful tool for large-scale genetic transformation and analysis of gene function.
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Received: 12 February 2014
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