A catalog of human cDNA expression clones and its application to structural genomics.
We describe here a systematic approach to the identification of human proteins and protein fragments that can be expressed as soluble proteins in Escherichia coli. A cDNA expression library of 10,825 clones was screened by small-scale expression and purification and 2,746 clones were identified. Sequence and protein-expression data were entered into a public database. A set of 163 clones was selected for structural analysis and 17 proteins were prepared for crystallization, leading to three new structures.
Büssow
Konrad
K
Protein Structure Factory, Heubnerweg 6, 14059 Berlin, Germany. buessow@molgen.mpg.de
Quedenau
Claudia
C
Sievert
Volker
V
Tischer
Janett
J
Scheich
Christoph
C
Seitz
Harald
H
Hieke
Brigitte
B
Niesen
Frank H
FH
Götz
Frank
F
Harttig
Ulrich
U
Lehrach
Hans
H
D002377:Catalogs as Topic; D003001:Cloning, Molecular / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods; D018076:DNA, Complementary / Q000096:biosynthesis*; D030541:Databases, Genetic; D015870:Gene Expression / Q000235:genetics; D015723:Gene Library; D023281:Genomics / Q000379:methods*; D006801:Humans; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D011237:Predictive Value of Tests; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods; D012995:Solubility
D018076:DNA, Complementary; D010446:Peptide Fragments; D011994:Recombinant Proteins