[Optimization of recombinant clumping factor of Staphylococcus aureus expression in Escherichia coli XLI-BLUE].
Experiments for optimization of the clfA gene expression in E. coli XLI-Blue cells were conducted. The reducing of the strain efficiency and the elimination of the plasmid during the cultivation of a producer have been shown. Several methods for raising the amount of plasmid-containing cells and increasing the yield of recombinant protein were proposed. Bacterial growth medium was selected to make this process more efficient. In such conditions after IPTG induction of the recombinant cells we have received high level of a soluble protein (about 30% of total cellular protein). The recombinant protein containing the His tag on N-terminus was purified on IDA-Sepharose column with high yield.
Ivanova
V D
VD
Taras Shevchenko National University, Kyiv, Ukraine. victdzani@yahoo.com
Pozur
V K
VK
D018929:Cell Culture Techniques / Q000379:methods; D003030:Coagulase / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
D003030:Coagulase; D011994:Recombinant Proteins