The alliance for cellular signaling plasmid collection: a flexible resource for protein localization studies and signaling pathway analysis.
Cellular responses to inputs that vary both temporally and spatially are determined by complex relationships between the components of cell signaling networks. Analysis of these relationships requires access to a wide range of experimental reagents and techniques, including the ability to express the protein components of the model cells in a variety of contexts. As part of the Alliance for Cellular Signaling, we developed a robust method for cloning large numbers of signaling ORFs into Gateway entry vectors, and we created a wide range of compatible expression platforms for proteomics applications. To date, we have generated over 3000 plasmids that are available to the scientific community via the American Type Culture Collection. We have established a website at www.signaling-gateway.org/data/plasmid/ that allows users to browse, search, and blast Alliance for Cellular Signaling plasmids. The collection primarily contains murine signaling ORFs with an emphasis on kinases and G protein signaling genes. Here we describe the cloning, databasing, and application of this proteomics resource for large scale subcellular localization screens in mammalian cell lines.
Zavzavadjian
Joelle R
JR
Alliance for Cellular Signaling, California Institute of Technology, CA 94305, USA.
Couture
Sam
S
Park
Wei Sun
WS
Whalen
James
J
Lyon
Stephen
S
Lee
Genie
G
Fung
Eileen
E
Mi
Qingli
Q
Liu
Jamie
J
Wall
Estelle
E
Santat
Leah
L
Dhandapani
Kavitha
K
Kivork
Christine
C
Driver
Adrienne
A
Zhu
Xiaocui
X
Chang
Mi Sook
MS
Randhawa
Baljinder
B
Gehrig
Elizabeth
E
Bryan
Heather
H
Verghese
Mary
M
Maer
Andreia
A
Saunders
Brian
B
Ning
Yuhong
Y
Subramaniam
Shankar
S
Meyer
Tobias
T
Simon
Melvin I
MI
O'Rourke
Nancy
N
Chandy
Grischa
G
Fraser
Iain D C
ID
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D016208:Databases, Factual; D051379:Mice; D016366:Open Reading Frames / Q000235:genetics; D010957:Plasmids; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics; D015398:Signal Transduction
D018076:DNA, Complementary; D011494:Protein Kinases