Appeal No. 2001-2500
Application No. 08/590,729
protein of interest, said sample incubated in a coupled
transcription/translation medium together with an aminoacyl
tRNA having fluorescent label;
(b) partially purifying said fluorescently labeled protein by
separating newly synthesized, fluorescently-labeled protein
from other fluorescent components within said sample;
(c) measuring the amount of protein synthesized;
(d) determining fluorescence of newly synthesized protein; and
(e) determining the biological activity of the newly synthesized
protein.
2. The method of claim 1, wherein said flurosrescent label is on
the N-terminus of said protein.
The examiner relies on the following references:
Hildenbrand et al. (Hildenbrand), “Sugar transport by the bacterial
phosphotransferase system,” The Journal of Biological Chemistry, Vol. 257,
No. 23, pp. 14518-14525 (1952)
Stryer, “Protein synthesis in bacteria is initiated by formylmethionyl transfer rna,”
Biochemistry, pp. 752-758 (1988)
Picking et al. (Picking), “Fluorescence characterization of the environment
encountered by nascent polyalanine and polyserine as they exit Escherichia coli
ribosomes during translation,” Biochemistry, Vol. 31, pp. 2368-2375 (1992)
Kudlicki et al. (Kudlicki), ”High efficiency cell-free synthesis of proteins :
refinement of the coupled transcription/translation system,” Analytical
Biochemistry, Vol. 206, pp. 389-393 (1992)
Claims 1 and 5-9 stand rejected under 35 U.S.C. § 103 as obvious over the
combined disclosures of Kudlicki, Picking, and Hildenbrand.
Claims 2-4 stand rejected under 35 U.S.C. § 103 as obvious over the
combined disclosures of Kudlicki, Picking, Hildenbrand, and Stryer.
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