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Construction of Bacterial Phytase Gene in pEGFP-N3 Vectors and Its Expression in COS7 Cells
LIU Fang1，LIU Zhen1，CHENG Jia1，FU Gui-hong2，WANG Huan1，HUANG Ying1，ZHANG Jian-she1＊
(1. Department of Bioengineering and Environment Science, Changsha University , Changsha 410003, Hunan, China；2. College of Animal Science, Hunan Agriculture University, Changsha 410128 , Hunan, China )
Abstract: A bacterial phytase cDNA was amplified with regular PCR technique and it contains 1360 bp encoding 423 amino acids with protein molecular size approximately 48 KDa. The cDNA fragment was sub-cloned into pEGFP-N3 plasmid DNA at the cloning sizes of BamH1/Pst1 .The recombinant pEGFP/ aPPA2 plasmid DNA were transfected into mammalian COS7 cells. The recombinant plasmid was expressed in the mammalian cells and the fusion proteins exhibited high phytase activity. This study provides a new system to produce phytase with a potential for an application in related industry.
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