Construct for overexpressing recombinant protein, useful e.g. for producing therapeutic proteins, has the downstream ribosome binding region deleted or mutated

摘要

Construct (A) for expressing a gene (I) that encodes a recombinant protein (II), under control of the tryptophan operon (Ptrp) in a prokaryotic host cell has, directly downstream of the initiation codon, a fully defined sequence (S1) as given in the specification in which at least one nucleotide (nt) is mutated or deleted so as to allow overexpression of (II), is new. (S1) has formula AAAGCAATTTTCGTACTG and is derived from bacterial sequences that correspond to a ribosome-binding site (RBS). Downstream of (S1) is a multiple cloning site (MCS) into which (I) is inserted. Independent claims are also included for the following: (1) vector containing (A); (2) prokaryotic host cells transformed with the vector; and (3) preparation of (II) using (A).