Signal sequence trap: a cloning strategy for secreted proteins and type I membrane proteins.
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.
Tashiro
K
K
Department of Medical Chemistry, Kyoto University Faculty of Medicine, Japan.
Tada
H
H
Heilker
R
R
Shirozu
M
M
Nakano
T
T
Honjo
T
T
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
D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D016207:Cytokines; D021382:Protein Sorting Signals; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins