Effects of aphidicolin on retrovirus DNA synthesis in vivo |
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Authors: | K Hagino-Yamagishi K Kano Y Mano |
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Institution: | Department of Biochemistry and Microbiology Rutgers University, Cook College New Brunswick, New Jersey 08903 USA |
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Abstract: | Renaturation of green-fluorescent protein (A-GFP) was achieved for the first time following denaturation in guanidine-HCl or acid. Denaturation was accompanied by the concerted loss of visible fluorescence, alteration of absorption characteristics, and large negative deflection of CD signal in the far UV. Dialysis of a guanidine-denatured sample at pH 8 resulted in 64% renaturation (return to native absorption) and neutralization of an acid-denatured sample restored 90% of the native absorption. Renatured GFP is highly fluorescent and indistinguishable from native GFP with respect to the shape of excitation and emission spectra. Both native and denatured proteins exhibit resistance to trypsin hydrolysis and have identically broad pH and heat stability profiles, all of which suggest full renaturation. |
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Keywords: | MuLV murine leukemia virus MuSV murine sarcoma virus RDDPase RNA-dependent DNA polymerase BSA bovine serum albumin |
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