Selection of cDNAs using chromosome-specific genomic clones: application to human chromosome 13.
We have developed a general method for en masse isolation of cDNAs present in a normalized library by hybridization to arrayed chromosome-specific phage lambda clones; we have used this approach to initiate exon-mapping of human chromosome 13. An advantage of the simultaneous isolation of cDNA/lambda pairs is that it allows cytogenetic assignment of a bona fide genomic clone by in situ hybridization, which also verifies that the corresponding cDNA or a homologous expressed sequence resides on chromosome 13. This information is enriched by partial sequencing of a selected cDNA from both ends. The sequence of the 3' noncoding region provides an 'identifier' that is used to develop STSs, while the sequence from the 5' end, often corresponding to a coding region, is used for homology searches in databases that occasionally reveal gene functions.
Department of Psychiatry, Columbia University, New York, NY 10032.
M F
MF
Bonaldo
M T
MT
Yu
P
P
Jelenc
S
S
Brown
L
L
Su
L
L
Lawton
L
L
Deaven
A
A
Efstratiadis
D
D
Warburton
M B
MB
Soares
D000595:Amino Acid Sequence; D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D002882:Chromosomes, Human, Pair 13; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006609:High Mobility Group Proteins / Q000235:genetics; D006801:Humans; D006822:Hybrid Cells; D008969:Molecular Sequence Data; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000235:genetics; D011494:Protein Kinases / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D016324:Sequence Tagged Sites; D014404:Tubulin / Q000235:genetics
D018076:DNA, Complementary; D006609:High Mobility Group Proteins; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D014404:Tubulin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D011494:Protein Kinases