RRC ID 48817
Author Shirozu M, Tada H, Tashiro K, Nakamura T, Lopez ND, Nazarea M, Hamada T, Sato T, Nakano T, Honjo T.
Title Characterization of novel secreted and membrane proteins isolated by the signal sequence trap method.
Journal Genomics
Abstract We recently described a method, called the signal sequence trap (SST) method, to clone cDNAs of secreted proteins and/or type I transmembrane proteins containing N-terminal signal sequences by using an epitope-tagging expression plasmid vector. In this paper we describe the summary of a large-scale screening of approximately 5900 clones of an SST cDNA library constructed from mouse bone marrow stromal cell line ST-2 cells. Of 26 positive clones obtained and sequenced, 11 clones appeared to contain authentic signal sequences. Five of the clones corresponded to the 5' ends of the cDNA of known genes containing N-terminal signal sequences. The full-length cDNA clones of the 6 other unknown clones were isolated and sequenced. One clone, termed SDF3, encoded a mouse counterpart of human pigment epithelium-derived factor. Another clone, termed SDR1, had considerable homology with basigin, a member of the immunoglobulin superfamily. A third clone, termed SDF5, had partial homology with a Drosophila tissue polarity gene frizzled (fz) and its rat homologues, fz-1 and fz-2. The other three clones had no significant homology with sequences in the databases. These results indicate that the SST method is effective and useful for the isolation of secreted and membrane proteins without knowledge of their functions.
Volume 37(3)
Pages 273-80
Published 1996-11-1
DOI 10.1006/geno.1996.0560
PII S0888-7543(96)90560-1
PMID 8938438
MeSH Amino Acid Sequence Animals Antigens, CD* Antigens, Neoplasm* Antigens, Surface* Avian Proteins* Basigin Blood Proteins* Bone Marrow / chemistry Cloning, Molecular / methods* DNA, Complementary / genetics* Drosophila Proteins* Drosophila melanogaster / genetics Frizzled Receptors Gene Library Humans Male Membrane Glycoproteins / chemistry Membrane Proteins / chemistry Membrane Proteins / genetics Membrane Proteins / isolation & purification* Membrane Proteins / secretion Mice Molecular Sequence Data Rats Receptors, G-Protein-Coupled Sequence Alignment Sequence Homology, Amino Acid Signal Transduction* Species Specificity
IF 2.91
Times Cited 66
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY GENETICS & HEREDITY
Resource
DNA material mouse SDF-5 cDNA (RDB06782) mouse SDR-1 alpha cDNA (RDB06783) mouse SDR-1 beta cDNA (RDB06784).