Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 147-157 |
| Number of pages | 11 |
| Journal | Journal of Structural and Functional Genomics |
| Volume | 5 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - 2004 |
| Externally published | Yes |
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Keywords
- Cloning
- Crystallization
- Expression
- High throughput
- TB structural genomics
ASJC Scopus subject areas
- Genetics
- Structural Biology
- Biochemistry
Cite this
Laboratory scale structural genomics. / Segelke, Brent W.; Schafer, Johana; Coleman, Matthew A; Lekin, Tim P.; Toppani, Dominique; Skowronek, Krzysztof J.; Kantardjieff, Katherine A.; Rupp, Bernhard.
In: Journal of Structural and Functional Genomics, Vol. 5, No. 1-2, 2004, p. 147-157.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Laboratory scale structural genomics
AU - Segelke, Brent W.
AU - Schafer, Johana
AU - Coleman, Matthew A
AU - Lekin, Tim P.
AU - Toppani, Dominique
AU - Skowronek, Krzysztof J.
AU - Kantardjieff, Katherine A.
AU - Rupp, Bernhard
PY - 2004
Y1 - 2004
N2 - 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.
AB - 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.
KW - Cloning
KW - Crystallization
KW - Expression
KW - High throughput
KW - TB structural genomics
UR - http://www.scopus.com/inward/record.url?scp=3543110884&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=3543110884&partnerID=8YFLogxK
U2 - 10.1023/B:JSFG.0000029193.82120.d1
DO - 10.1023/B:JSFG.0000029193.82120.d1
M3 - Article
C2 - 15263853
AN - SCOPUS:3543110884
VL - 5
SP - 147
EP - 157
JO - Journal of Structural and Functional Genomics
JF - Journal of Structural and Functional Genomics
SN - 1345-711X
IS - 1-2
ER -