Using a bioinformatical approach, a hybrid gene encoding the Fc-binding domains of both SPA and SPG (1 269 bp) was designed and synthesized. The sequence was designed by substitution of high-usage codons for low-usage ones of Escherichia coli. A total of 64 oligonucleotides were assembled in a single-step, in which T4 DNA ligase ligated DNA fragments from a pool of overlapping oligonucleotides. The synthetic gene was cloned to pSK vector. After sequencing and remending, the gene was expressed using the expression vector pET-28a-c(+). A band of 47 ku was detected with Western blot analysis. The protein so called MproteinAG has been covalently conjugated to the carboxyl-terminated magnetic particles. With the MproteinAG magnetic particles,IgG from blood serium of rabbit, pig,dog,mouse, human, macaque, rat, cat, sheep, cattle, horse can be purificated in one hour.
MENG Zhao-Hui, LIN Bing, XIE Yue-Hui, ZHANG Ke-Qin. Cloning and Expression of Fused Fc-binding Protein(SPA-SPG) and Its Application in Purification of IgG[J]. Progress in Biochemistry and Biophysics,2004,31(2):146-149
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