The isolation of structural genes from libraries of eucaryotic DNA

We present a procedure for eucaryotic structural gene isolation which involves the construction and screening of cloned libraries of genomic DNA. Large random DNA fragments are joined to phage lambda vectors by using synthetic DNA linkers. The recombinant molecules are packaged into viable phage par...

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Veröffentlicht in:Cell 1978-10, Vol.15 (2), p.687-701
Hauptverfasser: Maniatis, Tom, Hardison, Ross C., Lacy, Elizabeth, Lauer, Joyce, O'Connell, Catherine, Quon, Diana, Sim, Gek Kee, Efstratiadis, Argiris
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Sprache:eng
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Zusammenfassung:We present a procedure for eucaryotic structural gene isolation which involves the construction and screening of cloned libraries of genomic DNA. Large random DNA fragments are joined to phage lambda vectors by using synthetic DNA linkers. The recombinant molecules are packaged into viable phage particles in vitro and amplified to establish a permanent library. We isolated structural genes together with their associated sequences from three libraries constructed from Drosophila, silkmoth and rabbit genomic DNA. In particular, we obtained a large number of phage recombinants bearing the chorion gene sequence from the silkmoth library and several independent clones of β-globin genes from the rabbit library. Restriction mapping and hybridization studies reveal the presence of closely linked β-globin genes.
ISSN:0092-8674
1097-4172
DOI:10.1016/0092-8674(78)90036-3