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《Progress in Veterinary Medicine》 2018-05
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Cloning and Prokaryotic Expression of Penton Gene of Fowl Adenovirus Group Ⅰ Serotype Four

GAO Hui;ZHANG Meng-he;FENG Ru;TANG Ya-fang;ZHANG Hao;CHEN Cheng;LV Tao;WANG Cheng-bao;YANG Yan-tao;College of Veterinary Medicine,Northwest A&F University;Vocational and Technical Education Center;College of Life Sciences,Northwest A&F University;  
This study aimed to clone and prokaryotically express the Penton gene of fowl adenovirus group I serotype 4(FAdV-4),to obtain a high degree of accumulation of soluble Penton protein products.A pair of primers were designed according to the sequences of the Penton gene published in the GenBank.The PCR amplification and DNA sequencing analysis were performed using the total genomic virus DNA extracted from the purified FAdV-4 virus preserved in our laboratory as the template.The Penton gene was cloned into the pCold-SUMO vector,and then the protein was induced and expressed.The sequence analysis showed that the isolate was classified as FAdV-4,named SX17 strain(GenBank accession number:MF592716).And it belongs to the new mutant strains of domestic epidemic strains in recent years.SDSPAGE analysis and Western blot analysis showed that the Penton protein of SX17 strain was expressed as soluble protein.The expression and purification of the Penton protein showed to be with the high qualities,and the protein purity could be up to 80%-90% after separation.To our knowledge,this is the first report on successfully expressing and purifying soluble Penton protein of FAdV-4 in E.coli.The purified protein was proved to be with high purity.These data will be very helpful for the further study on the biological activity of FAdV-4 Penton protein and the following antibody preparation.
【Fund】: 国家自然科学基金项目(31302103);; 陕西省自然科学基础研究计划项目(2016JQ3010)
【CateGory Index】: S852.65
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