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Isolation of cDNA sequences coding for a part of human tissueplasminogen activator.
Edlund T
,
Ny T
,
Rånby M
,
Hedén LO
,
Palm G
,
Holmgren E
,
Josephson S
.
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We have isolated a cDNA sequence coding for a part of human tissueplasminogen activator. mRNA coding for tissueplasminogen 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 tissueplasminogen 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 tissueplasminogen activator.
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