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Isolation of cDNA clones for the human transferrin receptor.
Author(s) -
Schneider C.,
Kurkinen M.,
Greaves M.
Publication year - 1983
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1983.tb01732.x
Subject(s) - polysome , cdna library , biology , complementary dna , microbiology and biotechnology , transferrin receptor , sepharose , messenger rna , coding region , rna , receptor , gene , biochemistry , ribosome , enzyme
A cDNA clone bank containing 30 000 clones was constructed from sucrose gradient‐fractionated mRNA from human placenta. mRNA coding for transferrin receptor (TR) was enriched by polysome immuno‐adsorbed chromatography with monospecific rabbit IgG and protein‐A Sepharose. The library was screened for hybridisation to 32P‐labelled cDNA synthesised from immunoselected TR mRNA and from poly(A)+ RNA of the polysome fraction that failed to bind to protein‐A Sepharose. Plasmids isolated from colonies showing hybridisation only to the probe made from immunoselected mRNA were then subjected to hybrid selection. Two clones, pTR‐48 and pTR‐67, were able to hybridise the mRNA coding for the TR.