A signal sequence trap based on a constitutively active cytokine receptor.

Abstract:

:Targeting of secreted and cell-surface proteins to the cell membrane is mediated by a short hydrophobic stretch of amino acids, termed the signal sequence. We have developed a method that detects signal sequences in cDNA fragments based on their ability to redirect a constitutively active mutant of a cytokine receptor to the cell surface, thereby permitting interleukin-3 (IL-3)-independent growth of Ba/F3 cells. Retrovirus-mediated expression of the fusions in IL-3-dependent cells was followed by selection of clones for growth in the absence of IL-3. Infection of cells with 5x10(6) viral particles in a pilot experiment led to the isolation of 150 known and 48 novel cDNA clones, and all the known cDNA clones were found to encode secreted and cell-surface proteins. In addition, we isolated type II membrane proteins, which have not been detected by existing signal sequence trap strategies.

journal_name

Nat Biotechnol

journal_title

Nature biotechnology

authors

Kojima T,Kitamura T

doi

10.1038/8666

subject

Has Abstract

pub_date

1999-05-01 00:00:00

pages

487-90

issue

5

eissn

1087-0156

issn

1546-1696

journal_volume

17

pub_type

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