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Abstract
+ Q* Z2 E# C) X: l" d- ^" z5 e: mMultiple genetic modifications in pigs can essentially benefit research on agriculture, human disease and xenotransplantation.2 f. `# T. V. X0 x+ x+ W
Most multi-transgenic pigs have been produced by complex and time-consuming breeding programs using multiple" J/ Y! y) {( l! t$ @' K
single-transgenic pigs. This study explored the feasibility of producing multi-transgenic pigs using the viral 2A peptide in
/ }6 ^1 x' n8 T/ r0 R" B4 \. xthe light of previous research indicating that it can be utilized for multi-gene transfer in gene therapy and somatic cell
2 g' H" t' }: E9 D9 F1 r6 y9 x& Freprogramming. A 2A peptide-based double-promoter expression vector that mediated the expression of four fluorescent6 ?. a. [! f4 D1 p
proteins was constructed and transfected into primary porcine fetal fibroblasts. Cell colonies (54.3%) formed under G418
% W9 G+ z: q% p7 Z/ J* o# zselection co-expressed the four fluorescent proteins at uniformly high levels. The reconstructed embryos, which were
9 y* D% l' C) E2 ~% F6 ^6 Kobtained by somatic cell nuclear transfer and confirmed to express the four fluorescent proteins evenly, were transplanted. w# J- b7 a, b+ g& s2 t5 q; Z
into seven recipient gilts. Eleven piglets were delivered by two gilts, and seven of them co-expressed the four fluorescent
& v3 F( M+ D$ V M1 \3 I, sproteins at equivalently high levels in various tissues. The fluorescence intensities were directly observed at the nose, hoof, W! E& Y" a0 b1 S% i
and tongue using goggles. The results suggest that the strategy of combining the 2A peptide and double promoters" s7 z" A1 G6 v2 t
efficiently mediates the co-expression of the four fluorescent proteins in pigs and is hence a promising methodology to
' K5 q+ X5 E( p( B P; k3 Mgenerate multi-transgenic pigs by a single nuclear transfer. |
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发表于 2011-5-25 13:08
发表于 2011-5-25 13:14
