Rapid genomic scanning methods are required to identify expressed sequences and we report an efficient, sensitive and specific approach which relies upon hybridization of an amplified, labeled cDNA library to digested cosmid DNA. We identified expressed sequences within a cosmid in the glycerol kinase (GK) "critical region" of Xp21 that had impressive similarity to prokaryotic GKs. We used this genomic sequence information to clone the human hepatic GK cDNA. Independent confirmation of the identity of this gene was obtained by functional complementation of GK deficient E. coli mutants with a construct containing the complete human X-linked GK coding sequence.
Pubmed ID: 8401584 RIS Download
Mesh terms: Amino Acid Sequence | Bacillus subtilis | Base Sequence | Chromosome Mapping | Cloning, Molecular | Cosmids | Escherichia coli | Gene Library | Glycerol Kinase | Humans | Liver | Molecular Sequence Data | Oligodeoxyribonucleotides | Polymerase Chain Reaction | Sequence Homology, Amino Acid | X Chromosome
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