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| Structure Analysis of P8 Protein of Rice Gall Dwarf Virus and it's Expression in E.coli |
| GAO Fang-Luan1, FAN Guo-Cheng2, SHEN Jian-Guo2, XIE Li-Yan2, LIN Qi-Ying2, WU Zu-Jian1, XIE Lian-Hui2 |
1.Key Lab for Plant Virology of Fujian Province,Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou350002,China
2.Research Institute of Pomology, Fujian Academy of Agricultural Sciences, Fuzhou350013,China
3.Fujian ExitEntry Inspection and Quarantine Bureau, Fuzhou350001,China |
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Abstract To elucidate structural characters and expression characters of RGDV P8 protein in E.coli, bioinformatics analysis were performed by using biology software and analysis tools online.P8 gene was successfully amplified by RT-PCR and inserted to pGEX-4T-1 expression vector. The protein was then expressed in E.coli BL21(DE3) with IPTG induction. The antiserum was produced by immunizing rabbit with fusion protein, and the specificity of antiserum was evaluated by detecting RGDV, RDV, RSV, RRSV. Analysis showed that P8 protein had a typical transmembrane helices region, and a Phytoreo P8 domain shared with Phytoreo P8 family. SDSPAGE and Western blotting analysis showed that molecular weight of P8 fusion protein was about 73 kDa. P8 protein was efficiently expressed in the form of inclusion body. The antiserum with high titer and specificity provides the basis for detecting RGDV and studying function of P8 protein.
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Received: 19 March 2009
Published: 28 July 2009
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