Laboratory scale structural genomics.
At Lawrence Livermore National Laboratory, the development of the TB structural genomics consortium crystallization facility has paralleled several local proteomics research efforts that have grown out of gene expression microarray and comparative genomics studies. Collective experience gathered from TB consortium labs and other centers involved in the NIH-NIGMS protein structure initiative allows us to explore the possibilities and challenges of pursuing structural genomics on an academic laboratory scale. We discuss our procedures and protocols for genomic targeting approaches, primer design, cloning, small scale expression screening, scale-up and purification, through to automated crystallization screening and data collection. The procedures are carried out by a small group using a combination of traditional approaches, innovative molecular biochemistry approaches, software automation, and a modest investment in robotic equipment.
Segelke
Brent W
BW
Macromolecular Crystallography and Structural Genomics Group, Biology and Biotechnology Research Program, P.O. Box 808, Lawrence Livermore National Laboratory, Livermore, CA 94551, USA.
Schafer
Johana
J
Coleman
Matthew A
MA
Lekin
Tim P
TP
Toppani
Dominique
D
Skowronek
Krzysztof J
KJ
Kantardjieff
Katherine A
KA
Rupp
Bernhard
B
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D023281:Genomics / Q000379:methods*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012371:Robotics
D011994:Recombinant Proteins