| 主权项 |
1. An expression cassette comprising a nucleotide sequence with the operably linked nucleic acid sequences:
5′ Pr-(TIS)D(IBFP1)E-(CL1)G-ORF-[CL2-ORF]L-(CL3)M-(IBFP2)Q-(SSC)R-(CL4)T-(Ft)W-(Tr)X-3′wherein
the Pr is a promoter sequence, the TIS encodes a translation initiation sequence, the IBFP1 encodes a first inclusion body fusion partner comprising an amino acid sequence of any of SEQ ID NOS: 7-14 or a variant thereof, wherein the variant has 75% or greater sequence homology to at least one of SEQ ID NOS: 7-14 and wherein the variant includes a hydrophobic core consisting of residues 22-29 of SEQ ID NO:130, the CL1 encodes a first cleavable peptide linker, the ORFs encode a preselected polypeptide, the CL2 encodes a second cleavable peptide linker, the CL3 encodes a third cleavable peptide linker, the IBFP2 encodes a second inclusion body fusion partner comprising an amino acid sequence of any of SEQ ID NOS: 7-14 or a variant thereof, wherein the variant has 75% or greater sequence homology to at least one of SEQ ID NOS: 7-14 and wherein the variant includes the hydrophobic core consisting of residues 22-29 of SEQ ID NO:130, the SSC is a suppressible stop codon, the CL4 encodes a fourth cleavable peptide linker, the Ft encodes a fusion tag, and the Tr is a transcription terminator sequence, wherein each of D or X is independently 0 or an integer of 1 to 4, wherein R is 0 or an integer of 1 to 2, wherein each of E, G, L, M, Q, T or W is independently 0 or an integer of 1 to 20, wherein either one or both of a IBFP1 or a IBFP2 is present, and wherein expression of the nucleic acid sequence produces a tandem polypeptide that forms an inclusion body when expressed in a cell, wherein at least one of a CL1, a CL2, a CL3 or a CL4 is present. |
