A genome annotation-driven approach to cloning the human ORFeome.
We have developed a systematic approach to generating cDNA clones containing full-length open reading frames (ORFs), exploiting knowledge of gene structure from genomic sequence. Each ORF was amplified by PCR from a pool of primary cDNAs, cloned and confirmed by sequencing. We obtained clones representing 70% of genes on human chromosome 22, whereas searching available cDNA clone collections found at best 48% from a single collection and 60% for all collections combined.
The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SA, UK.
John E
JE
Collins
Charmain L
CL
Wright
Carol A
CA
Edwards
Matthew P
MP
Davis
James A
JA
Grinham
Charlotte G
CG
Cole
Melanie E
ME
Goward
Begoña
B
Aguado
Meera
M
Mallya
Younes
Y
Mokrab
Elizabeth J
EJ
Huckle
David M
DM
Beare
Ian
I
Dunham
D002892:Chromosomes, Human, Pair 22 / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D019295:Computational Biology; D018076:DNA, Complementary / Q000235:genetics; D030541:Databases, Genetic; D015894:Genome, Human; D023281:Genomics / Q000379:methods*; D006801:Humans; D016366:Open Reading Frames / Q000235:genetics*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D020543:Proteome / Q000235:genetics*; D012107:Research Design; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D020543:Proteome