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Abstract
" Z- S' A; T2 DMultiple genetic modifications in pigs can essentially benefit research on agriculture, human disease and xenotransplantation.
! C/ I. E1 D+ Y$ W) C# fMost multi-transgenic pigs have been produced by complex and time-consuming breeding programs using multiple) [% q% V7 J& |1 L" v- G( }" O
single-transgenic pigs. This study explored the feasibility of producing multi-transgenic pigs using the viral 2A peptide in
/ z. o, d3 Z/ {: u. l: k6 gthe light of previous research indicating that it can be utilized for multi-gene transfer in gene therapy and somatic cell
6 j, W! O. p2 Ureprogramming. A 2A peptide-based double-promoter expression vector that mediated the expression of four fluorescent
: O4 W2 r% {: h! \5 k Q6 S( Wproteins was constructed and transfected into primary porcine fetal fibroblasts. Cell colonies (54.3%) formed under G418
. b3 m+ u; ?$ k, W" _) ]selection co-expressed the four fluorescent proteins at uniformly high levels. The reconstructed embryos, which were
" M- M- h% e5 b/ Y( ^5 V: Y3 kobtained by somatic cell nuclear transfer and confirmed to express the four fluorescent proteins evenly, were transplanted
1 o0 c* g5 f0 f, Tinto seven recipient gilts. Eleven piglets were delivered by two gilts, and seven of them co-expressed the four fluorescent
3 b! y4 Q" X% K3 u) `proteins at equivalently high levels in various tissues. The fluorescence intensities were directly observed at the nose, hoof# R2 Q% I: y1 O( G
and tongue using goggles. The results suggest that the strategy of combining the 2A peptide and double promoters
. a( }# T# {! F+ ]efficiently mediates the co-expression of the four fluorescent proteins in pigs and is hence a promising methodology to
" W+ ^; |* S+ U" n9 e! B; pgenerate 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
