›› 2005, Vol. 48 ›› Issue (1): 13-17.
• RESEARCH PAPERS • Previous Articles Next Articles
GAO Bo1,LIU Zhi-Gang1,2*,XING Miao1,2,XU Hong2, LUO Shi-Wen2, LAI Ren2
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Abstract: Using the Cr PI clone from the λEXcell library as a template, the cDNA fragments were first generated by PCR techniques and then ligated into T vector. After being confirmed by DNA sequencing, the cDNA encoding the American cockroach Cr PI allergen was subcloned into pGEX-5X-1 and expressed as GSTfusion protein in the form of inclusion bodies. After being dissolved in 6 mol/L guanidine hydrochloride and renatured with a simple dilution method, the proteins of target were purified to above 90% purity by affinity chromatography with Glutathione Sepharose 4B. Tested with sera from subjects allergic to cockroach, the recombinant allergen was shown to possess good IgEbinding activity as determined by Western blotting.
Key words: American cockroach, Periplaneta americana, recombinant allergen Cr PI, protein expression, affinity chromatography, Western blotting
GAO Bo1,LIU Zhi-Gang1,2*,XING Miao1,2,XU Hong2, LUO Shi-Wen2, LAI Ren2. Expression, purification and characterization of the American cockroach Cr PI allergen[J]., 2005, 48(1): 13-17.
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