Signal sequence trap: a cloning strategy for secreted proteins and type I membrane proteins

Science. 1993 Jul 30;261(5121):600-3. doi: 10.1126/science.8342023.

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.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line
  • Chemokine CXCL12
  • Chemokines, CXC*
  • Cloning, Molecular / methods*
  • Cytokines / chemistry
  • Cytokines / genetics*
  • Genetic Vectors
  • Molecular Sequence Data
  • Open Reading Frames
  • Protein Sorting Signals / chemistry
  • Protein Sorting Signals / genetics*
  • Protein Sorting Signals / metabolism
  • Receptors, Interleukin-2 / genetics
  • Recombinant Fusion Proteins / biosynthesis
  • Transfection

Substances

  • Chemokine CXCL12
  • Chemokines, CXC
  • Cytokines
  • Protein Sorting Signals
  • Receptors, Interleukin-2
  • Recombinant Fusion Proteins

Associated data

  • GENBANK/L12029
  • GENBANK/L12030