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《China Biotechnology》 2008-06
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Overexpression and Down-stream Technology of Recombinant Sus scrofa Interferon alpha in Escherichia coli

LI Wen-hui1 LIU Di2 WANG Jun-wei3 XU Dong-mei4 YANG Xiu-qin1 (1 Animal Science Technical College Northeast Agricultural University,Harbin 150030,China) (2 Heilongjiang Academy of Agricultural Sciences,Harbin 150086,China 3 Northeast Agricultural University,Harbin 150030,China) (4 Bio-technology Company of Harbin Pharmaceutical Co., Ltd,Harbin 150076,China)  
A 501bp sequence of Sus scrofa IFN-α gene and Susscrota domestica of changbai IFN-α gene was amplified by normal polymerase chain reaction (PCR) from their genome whose homology are more than 98.4% ,compared with nucleotide sequence published on NCBI .Referring to Sus scrofa IFN-α gene sequence,a new sequence was synthesized after codon bias and mRNA structure optimizaed .Then ,this sequence was inserted in pWL vector and the recombinant plasmids were transformed into the E.coli DH-5α.After fermentation,the expression level of recombinant protein are 25% by SDS-PAGE analysis.The biologic activity was 4.45×106 IU/mg by VSV-WISH system analysis after protein refolding and purification.
【CateGory Index】: Q786
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