Cloning and Expression of the Gene for Pro-urokinase in Escherichia coli

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An Escherichia coli library containing cDNA sequences derived from mRNA of human pharnygeal carcinoma cells was constructed using the plasmid pBR322. The library was screened with a set of 8 deoxyoligonucleotides, 14 bases in length, which collectively code for the amino acid sequence …Met-Tyr-Asn-Asp-Pro… found in human urokinase (u-PA). A plasmid containing an 1700 base pair cDNA insert coding for the carboxy terminal sequence of u-PA was identified. The 5′ region was obtained by priming poly(A) mRNA with a DNA fragment derived from the initial clone. An expression vector was constructed which directs the synthesis of pro u-PA in E. coli.

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Holmes, W., Pennica, D., Blaber, M. et al. Cloning and Expression of the Gene for Pro-urokinase in Escherichia coli. Nat Biotechnol 3, 923–929 (1985) doi:10.1038/nbt1085-923

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