New vector construct comprising a DNA sequence that codes a polypeptide with or without a signal sequence and a DNA sequence that codes a cleavage site, for expression and secretion of polypeptides in filamentous fungi

摘要

Vector construct for the expression and secretion of polypeptides in filamentous fungi, comprising (along 5'3'-direction in functional linkage): optionally a promoter, which can be preceded by an enhancer; at least a DNA sequence, which codes a first polypeptide and may or may not contain a signal sequence; optionally a DNA sequence, which codes a 2A-cleavage site or codes a sequence derived from it, optionally together with a preceding DNA sequence that codes a proteolytic cleavage site or optionally a DNA sequence that codes a proteolytic cleavage site; and a terminator, is new. Vector construct for the expression and secretion of polypeptides in filamentous fungi, comprising (along the 5'3'-direction in functional linkage): optionally a promoter, which can be preceded by an enhancer; at least a DNA sequence, which codes a first polypeptide and may or may not contain a signal sequence; optionally a DNA sequence, which codes a 2A-cleavage site or codes a sequence derived from it, optionally together with a preceding DNA sequence that codes a proteolytic cleavage site or optionally a DNA sequence that codes a proteolytic cleavage site; a DNA sequence that codes a second polypeptide with a signal sequence; optionally an additional DNA sequence, which codes a 2A-cleavage site or a sequence that is derived from it, optionally together with a preceding DNA sequence that codes a proteolytic cleavage site or optionally a DNA sequence, which codes a proteolytic cleavage site; optionally a DNA sequence that codes a third polypeptide with a signal sequence; optionally an additional DNA sequence, which codes a 2A-cleavage site or a sequence that is derived from it, optionally together with a preceding DNA sequence that codes a proteolytic cleavage site or optionally a DNA sequence, which codes a proteolytic cleavage site; optionally a DNA sequence that codes a fourth polypeptide with a signal sequence; and a terminator, where in the construct at least a 2A-cleavage site or a sequence that is derived from it is present, is new. Independent claims are included for: (1) a recombinant polypeptide produced through a cell that is transformed with the vector construct; (2) a host cell transformed with the vector construct; (3) producing one or more polypeptides in a filamentous fungi, comprising: (i) transforming a host cell with the vector constructs, (ii) culturing the transformed host cell from (i), under the conditions that are suitable for the expression and the secretion of the polypeptide(s) and (iii) isolation of the so-expressed polypeptide(s); and (4) use of a DNA sequence, which codes a 2A-cleavage site or a sequence that is derived from it, optionally together with a preceding DNA sequence that codes a proteolytic cleavage site, for producing an expression system for the expression and secretion of polypeptides in the filamentous fungi.