Cloning of cDNA coding for human tissue-type plasminogen activator and its expression in Escherichia coli
Harris, T.J.; Patel, T.; Marston, F.A.; Little, S.; Emtage, J.S.; Opdenakker, G.; Volckaert, G.; Rombauts, W.; Billiau, A.; De Somer, P.
Molecular Biology and Medicine 3(3): 279-292
1986
ISSN/ISBN: 0735-1313 PMID: 3090401 Document Number: 272004
cDNA clones of the mRNA coding for tissue-type plasminogen activator (t-PA) have been obtained and their nucleotide sequences compared to those reported previously. A gene coding for t-PA has been reconstructed and inserted into vectors for expression in prokaryotic cells. Relatively high levels of t-PA accumulated in inclusion bodies in Escherichia coli containing an optimized expression plasmid, but only a small proportion of the insoluble protein was recovered as active enzyme using a variety of solubilization procedures.