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| Application of Ribosomal Skipping Based Polycistronic Co-expression Technique in Arabidopsis thaliana |
| WU Li-Zhu*, AN Ye-Zhi*, ZHANG Jie,SUN Tian-Jie,WANG Dong-Mei** |
| College of Life Sciences, Hebei Agricultural University, Baoding 071001, China |
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Abstract 2A peptide is a novel ribosomal skipping based technique, which takes advantage of a cis-acting hydrolase element of a length of 18~22 amino acids with profiles of short structure and high protein segmentation efficiency. Therefore, it is increasingly used in polycistronic expression vectors. At present, it is widely used in yeast (Saccharomyces cerevisiae) and mammal cells, less in plants. In order to explore 2A peptide application in plants, the polycistronic co-expression vector pX6-2A with Porcine teschovirus-1 2A (P2A) and Thosea asigna virus 2A (T2A) linking 3 ORFs (open reading fragments) including a chimeric transcription activator gene XVE (X, the DNA-binding domain of the bacterial repressor LexA; V, the acidic trans-activating domain of VP16; E, the regulatory region of the human estrogen receptor.), GFP (green fluorescent protein) and Bar (bialaphos resistance) in series under the control of 35S promoter was constructed, and transformed into Arabidopsis thaliana, to obtain successful transgenic plants. Western blot results showed that these 3 proteins XVE, GFP and Bar expressed separately, indicated that both P2A and T2A peptide could transcribe, express and translate normally, and there was no interference with each other. It suggested that it could be an ideal way to express polycistronic in plants. These results indicated that P2A and T2A could do 'ribosome skipping' and separate proteins completely in plants. P2A and T2A were suitable for application in plants, which provides an important theoretical and referential significance for the popularization and application of 2A peptide in transgenic plants.
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Received: 21 February 2019
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Corresponding Authors:
dongmeiwang63@126.com
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