Construction and application of polyproteins

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Enzymatic production of a protein or polypeptide

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435 70, 4351723, 935 29, 935 47, 536 27, C12P 2100, C12P 2102, C12N 1500, C07H 1512

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active

047741805

ABSTRACT:
A method for constructing polyproteins which can perform multiple sequential activities. A DNA sequence is constructed using genetic engineering techniques to insert sequences encoding the desired proteins into a plasmid, in the correct order, following a single promoter element and before a single stop codon. The reading frames of the mRNA sequences are phased so that a polyprotein with the desired activities, in the required order, is produced.
Modified polyproteins can be produced by inserting or substituting amino acids into the mRNA sequence to create spaces between the individual proteins, to increase the stability of the total polyprotein, to change the spatial orientation of the individual proteins relative to each other and their substrates, and to modify the activity of the individual proteins.

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"Deletion of an Essential Gene in Escherichia coli by Site-Specific Recombination with Linear DNA Fragments", Jasin et al., Journal of Bacteriology, (Aug. 1984), pp. 783-786, vol. 159(2).

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