Isolation of cDNA Sequences Coding for a Part of Human Tissue Plasminogen Activator

We have isolated a cDNA sequence coding for a part of human tissue plasminogen activator. mRNA coding for tissue plasminogen activator was partially purified, copied into double-stranded cDNA, and cloned into Escherichia coli. Two sets of partially overlapping oligodeoxynucleotide mixtures correspon...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1983-01, Vol.80 (2), p.349-352
Main Authors: Edlund, Thomas, Ny, Tor, Ranby, Mats, Heden, Lars-Olof, Palm, Gunnar, Holmgren, Erik, Josephson, Staffan
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Animals
Base Sequence
Cell Line
Cell lines
Cloning, Molecular
Complementary DNA
DNA
DNA - isolation & purification
Female
Humans
Melanoma
Messenger RNA
Nucleotide sequences
Oligonucleotide probes
Oligonucleotides
Oocytes - metabolism
Plasmids
Plasminogen Activators - genetics
Protein Biosynthesis
RNA
RNA, Messenger - genetics
Xenopus
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Summary:We have isolated a cDNA sequence coding for a part of human tissue plasminogen activator. mRNA coding for tissue plasminogen activator was partially purified, copied into double-stranded cDNA, and cloned into Escherichia coli. Two sets of partially overlapping oligodeoxynucleotide mixtures corresponding to all possible coding sequences for a known portion of the tissue plasminogen activator gene were prepared. One set was used as a probe to screen cDNA containing bacterial clones and both were used as probes in hybridization against purified plasmid DNA. Of 4,200 bacterial clones examined, 1 carried a plasmid that hybridized to both sets of oligonucleotides. This plasmid contained a 370-base-pair cDNA insert, which was shown by nucleotide sequence analysis to code for the cleavage site region in the one-chain form of the human tissue plasminogen activator.
ISSN:0027-8424
1091-6490
1091-6490
DOI:10.1073/pnas.80.2.349