Construction of Plant Expression Vector with Maize γ-zein Gene and GFP Gene and Their Subcellular Localization
2 Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing, 100094, P.R. China
Author Correspondence author
Maize Genomics and Genetics, 2012, Vol. 3, No. 2 doi: 10.5376/mgg.2012.03.0002
Received: 29 Feb., 2012 Accepted: 30 May, 2012 Published: 01 Jun., 2012
Zhang et al., 2012, Construction of Plant Expression Vector with Maizeγ-zein gene and GFP gene and their Subcellular Localization, Maize Genomics and Genetics, Vol.3, No.2, 6-12 (doi: 10.5376/mgg.2012.03.0002)
The full length of the maize γ-zein gene amplified by PCR from PROK.TG1LK plasmid was ligated into the intermediate vector pMD18-T, to obtain the recombinant plasmid named pMD18-γ-zein. After the recombinant plasmid and the improved pCAMBIAI1302,respectively were digested by Nco I and Bgl II restriction endonucleases, the plant expression vector pCB-GFP-γ-zein with the gene of green fluorescent protein (GFP) driven by 35S was built following the procedures of recovery, ligation, transformation and identification. The fusion of GFP-γ-zein gene was integrated into the onion epidermal cells by using the Biolistic approach; the fusion gene was expressed in onion epidermal cell wall, nucleus and cytoplasm confirmed by confocal microscopy.
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