Expression and Bioactivity of Recombinant Human Lysozyme in the Milk of Transgenic Mice

Diagrammatic representation of (A) the pBC-hLY expression vector and (B) the pBC-sighLY expression vector. 2x globin insulator = 2 copies of chicken β-globin insulator; Pcasein = goat β-casein promoter; Casein E1–E2 = goat β-Casein exon1, intron1, part of exon 2 (before initial codon); hLY encoding region = human lysozyme encoding region; Casein E7–E8–E9 = goat β-Casein exon 7, intron 7, exon 8, intron 8, and exon 9; Casein 3genome DNA = goat β-casein 3′ genomic region; sigpeptide DNA sequence = bovine β-casein signal peptide DNA sequence.

Identification of (A) pBC-hLY and (B) pBC-sighLY transgenic mice by Southern blot. M = 1kb DNA ladder; 10, 5, 1 in A represent 10, 5, 1 copies of pBC-hLY plasmid, respectively; 5, 3, 1 in B represent 5, 3, 1 copies of pBC-sighLY plasmid, respectively; ck = wild type; I-1, I-2, I-3, I-4, I-5, I-6 indicate no. 1, 2, 3, 4, 5, 6 pBC-hLY transgenic mouse, respectively; II-1, II-2, II-3, II-4, II-5 indicate no. 1, 2, 3, 4, 5 pBC-hLY transgenic mouse, respectively.

Western blot analysis of the whey from (A) pBC-hLY and (B) pBC-sighLY female transgenic mice. ck = milk from wild type mouse; human = human milk; I-1, I-3, I-4 indicate milk from no. 1, 3, 4 pBC-hLY transgenic mouse, respectively; II-2, II-4, II-5 indicate milk from no. 2, 4, 5 pBC-sighLY mouse, respectively.