TY - JOUR
T1 - The isolation of structural genes from libraries of eucaryotic DNA
AU - Maniatis, Tom
AU - Hardison, Ross C.
AU - Lacy, Elizabeth
AU - Lauer, Joyce
AU - O'Connell, Catherine
AU - Quon, Diana
AU - Sim, Gek Kee
AU - Efstratiadis, Argiris
N1 - Funding Information:
Anderson for suggestions and discussions, and to N. Davidson for critical readings of the manuscript. We thank F. Blattner for providing Charon phage strains; N. Sternberg and L. Enquist for providing in vitro packaging strains; C. K. ltakura and R. Scheller for providing Eco RI linkers; J. Rosenberg for providing Eco RI methylase; R. Robinson for providing labeled single-copy Drosophila DNA tracer; and A. Cortenbach for preparing media and materials. We thank W. Bender and Hogness for discussions and for communicating unpublished data on the Drosophila library characterization, and F. C. Kafatos for the use of facilities and his support. We thank J. Maniatis for help with the figures. This work was supported by an NSF grant to F. C. Kafatos and T. M.; an NIH grant to F. C. Kafatos. an American Cancer Society grant to T. M. and funds from an NIH Biomedical Research Support grant to the California Institute of Technology. R. C. H. was supported by a fellowship from the Jane Coffin Childs Memorial Fund for Medical Research. T. M. is the recipient of a Rita Allen Foundation career development award.
PY - 1978/10
Y1 - 1978/10
N2 - 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.
AB - 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.
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U2 - 10.1016/0092-8674(78)90036-3
DO - 10.1016/0092-8674(78)90036-3
M3 - Article
C2 - 719759
AN - SCOPUS:0018117473
SN - 0092-8674
VL - 15
SP - 687
EP - 701
JO - Cell
JF - Cell
IS - 2
ER -