Proteomic Analysis of Tomato ( Lycopersicum esculentum var. cerasifarm ) Expressing the HBsAg Gene by 2-dimensional Diff
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ORIGINAL PAPER
Proteomic Analysis of Tomato (Lycopersicum esculentum var. cerasifarm ) Expressing the HBsAg Gene by 2-dimensional Difference Gel Electrophoresis Bin Guo & Wei He & Daochang Wu & Delu Che & Penghui Fan & Lingling Xu & Yahui Wei
Published online: 21 September 2013 # Springer Science+Business Media New York 2013
Abstract In a previous study, an HBsAg gene-bearing transgenic tomato line was made available and it exhibited notable physiological alterations compared with the non-transgenic tomato (control). In particular, leaves of the transgenic plants were fleshy and dark. We hypothesized that a change in leaf proteins of the transgenic plants account for the observed phenotypes. In this study, total protein content in leaves of the transgenic plants was analyzed by 2-dimensional difference gel electrophoresis. A total number of 700 protein spots were detected on silver-stained gels, of which 368 protein spots were matched between the control and sample gels. Among these matched proteins, the expression levels of 122 proteins in the transgenic plants were upregulated while those of the rest were downregulated. In addition, 25 abundant proteins (value ratio> 2.0) on silver-stained gels were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Sixteen differentially expressed proteins were identified, of which 13 were predicted to be involved in cell division, energy metabolism, protein synthesis and processing. The possible roles of these proteins in the transgenic tomato strain have been discussed. Taken together, our data indicate that significant alterations in protein expression occur in transgenic tomatoes Electronic supplementary material The online version of this article (doi:10.1007/s11130-013-0387-4) contains supplementary material, which is available to authorized users. B. Guo : W. He : D. Wu : D. Che : P. Fan : Y. Wei (*) Key Laboratory of Resource Biology and Biotechnology in Western China, Northwest University, Xi’an 710069, China e-mail: [email protected] B. Guo : W. He : D. Wu : D. Che : P. Fan : Y. Wei Ministry of Education; School of Life Science, Northwest University, Xi’an 710069, China L. Xu Department of Life Sciences, Xi’an University of Arts and Science, Xi’an 710065, China
bearing the HBsAg gene. Our findings will help broaden our knowledge of the mechanism by which exogenously expressed genes lead to phenotypic alterations in transgenic plants. Keywords Proteome map . Lycopersicon esculentum . 2-dimensional gel . Electrophoresis . Phenotype variance
Introduction Hepatitis B virus (HBV) infection is one of the most widespread viral infections of humans, causing acute and chronic hepatitis and hepatocellular carcinoma [1]. Vaccination is one of the best solutions to prevent HBV infection. At present, the development of “edible” vaccines based on transgenic plants is a promising direction in developing novel types of vaccines. Studies showed that HBsAg could be expressed in plants such as tobacco, potato and banana [2, 3], and partially pu
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