title: Signal sequence trap: a cloning strategy for secreted proteins and type I membrane proteins.
abstract: A method was developed to clone, without the use of specific functional assays, complementary DNAs (cDNAs) that carry specific amino-terminal signal sequences, such as those encoding intercellular signal-transducing molecules and receptors. The vector used in this system directed the cell surface expression of interleukin-2 receptor fusion proteins when inserts with signal sequences were cloned in-frame with the correct orientation. An expression cDNA library was constructed from a bone marrow stromal cell line, which contained 5' portion-enriched cDNAs (the average size was 400 base pairs). Two cDNAs that encoded putative cytokine molecules, stromal cell-derived factor-1 alpha (SDF-1 alpha) and SDF-1 beta, which belong to the intercrine-macrophage inflammatory protein superfamily, were cloned.
authors.affiliation: Department of Medical Chemistry, Kyoto University Faculty of Medicine, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Tashiro
authors.forename: H
authors.initials: H
authors.lastname: Tada
authors.forename: R
authors.initials: R
authors.lastname: Heilker
authors.forename: M
authors.initials: M
authors.lastname: Shirozu
authors.forename: T
authors.initials: T
authors.lastname: Nakano
authors.forename: T
authors.initials: T
authors.lastname: Honjo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D003001:Cloning, Molecular / Q000379:methods*; D016207:Cytokines / Q000737:chemistry / Q000235:genetics*; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D021382:Protein Sorting Signals / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015375:Receptors, Interleukin-2 / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection
chemical_list: D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D016207:Cytokines; D021382:Protein Sorting Signals; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins