Construction of Plant Expression Vector with Maize [gamma]-zein Gene and GFP Gene and Their Subcellular Localization

• Published in: Maize Genomics and Genetics Vol. 3; no. 2
• Main Authors: Zhang, Yu, Bai, Shiqie, Li, Cong, Li, Daxu, Deng, Yongchang, Wang, Yongxin
• Format: Journal Article
• Language: English
• Published: Richmond Sophia Publishing Group Inc 01.05.2012
• ISSN: 1925-1971
• 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.