Monomeric red fluorescent proteins
AssigneesThe University of Chicago, University of Washington
InventorsBenjamin S. Glick, Brooke Bevis, Daniel E. Strongin, David Baker, Michelle Scalley-Kim
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Abstract
Disclosed are sequences encoding monomeric variants of DsRed fluorescent proteins and methods of use.
Claims7
An isolated polynucleotide encoding a variant polypeptide of wild-type DsRed of SEQ ID NO:1 or a variant of the rapidly maturing Discosoma red fluorescent protein T4 (DsRed.T4) of SEQ ID NO:3, the variant polypeptide having reduced oligomerization relative wild-type DsRed, the variant polypeptide comprising the amino acid substitutions K83M, K163H, and Y193H, the variant polypeptide further comprising at least one amino acid substitution selected from the group consisting of E26Y, K92T, V96S, T106E, T108Q, 1125K, S131A, 1180V, and M182K.
An isolated polynucleotide encoding a variant polypeptide of the rapidly maturing Discosoma red fluorescent protein T4 (DsRed.T4) of SEQ ID NO:3, the variant polypeptide having reduced oligomerization relative wild-type DsRed, wherein the variant matures more rapidly that wild-type DsRed, the variant polypeptide comprising:
- at least one amino acid substitution selected from the group consisting of K83M, K83L, K163Q, K163M, K163H, and Y193H;
- at least three amino acid substitutions selected from the group consisting of E26Y, K92T, V96S, T106E, T108Q, 1125K, S131A, 1180V, and M182K;
- at least three amino acid substitutions selected from the group consisting of R149K, R153Q, H162S, L174T, E176D, Y192N, R216H, H222S, L223G, and F224S;
- and
- at least one amino acid substitution selected from the group consisting of V71A, C117T, S175C, S179T, S203N, and G219A.
A variant polypeptide of DsRed.T4 (SEQ ID NO:3) comprising:
- at least one amino acid substitution selected from the group consisting of K83M, K83L, K163Q, K163M, K163H, and Y193H;
- at least three amino acid substitutions selected from the group consisting of E26Y, K92T, V96S, T106E, T108Q, 1125K, S131A, 1180V, and M182K;
- and
- at least three amino acid substitutions selected from the group consisting of R149K, R153Q, H162S, L174T, E176D, Y192N, R216H, H222S, L223G, and F224S
Dependencies are inferred from the claim text.