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[0371] For amplification of the genes for the human IgE heavy chain constant regions (IGHE) primers can be used containing an Ascl site (GAT CGG CGC GCC CAT CCG TCT TCC CCT TGA (SEQ ID NO:61)), an Sfil site, a 4xhis-tag (GAT CGG CCC AGC CGG CCT CAT TTA CCG GGA TTT ACA GAC AC (SEQ ID NO: 62)), and for the C H 2-4 domains primers containing an Ascl site (GAT CGG CGC GCC CAC CGT GAA GAT CTT AC (SEQ ID NO:63)), an Xbal site, and a 4xhis-tag (GAT CTC TAG ATC AAT GGT GGT GAT GTT TAC CGG GAT TTA CAG ACA CCG (SEQ ID NO: 64)) can be used

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LET7 H290N or LET7 W372F was PCR-amplified from pHBT -p35S :: LET7 H290N / LET7 W372F as a template and constructed into the pET28a-HIS-SUMO vector

Step 2: Support with Vitamin C