US 7,351,814 B2
Olfactory receptor expression libraries and methods of making and using them
Randall R. Reed, Baltimore, Md. (US); King-Wai Yau, Towson, Md. (US); and Dietmar Krautwurst, Bergholz Rebhrucke (Germany)
Assigned to The Johns Hopkins University, Baltimore, Md. (US)
Filed on Oct. 25, 2005, as Appl. No. 11/257,062.
Application 10/259430 is a division of application No. 09/465901, filed on Dec. 17, 1999, granted, now 6,492,143.
Application 11/257062 is a continuation of application No. 10/259430, filed on Sep. 30, 2002, abandoned.
Claims priority of provisional application 60/112605, filed on Dec. 17, 1998.
Prior Publication US 2006/0094047 A1, May 04, 2006
Int. Cl. C12N 15/12 (2006.01)
U.S. Cl. 536—23.5  [536/23.1] 8 Claims
 
1. A nucleic acid construct that encodes a hybrid seven-transmembrane receptor which facilitates the translocation of a seven-transmembrane receptor across a cell membrane, said construct comprising:
(i) a first nucleic acid sequence that encodes a first seven-transmembrane receptor polypeptide that is the rhodopsin translocation peptide contained in SEQ ID NO:3 or a nucleic acid sequence encoding a variant of said rhodopsin translocation peptide, wherein said variant is identical to said rhodopsin translocation peptide but for a single conservative amino acid substitution which substitutes a first amino residue in a first group for a second amino acid residue within the first group, wherein the first group is selected from the groups consisting of: 1) Alanine (A), Serine (S), Threonine (T); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); and
(ii) a second nucleic acid sequence that encodes a second seven-transmembrane receptor polypeptide, wherein said nucleic acid construct when expressed in a cell results in a hybrid seven-transmembrane receptor polypeptide comprising the rhodopsin translocation peptide or said variant, and the second seven-transmembrane receptor polypeptide, wherein the rhodopsin translocation peptide or said variant directs the translocation of the second seven-transmembrane receptor polypeptide across the cell membrane, wherein said variant translocates the seven-transmembrane receptor polypeptide as efficiently as the rhodopsin translocation polypeptide.