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Abstract. m% Q/ [) m1 ], Z
Multiple genetic modifications in pigs can essentially benefit research on agriculture, human disease and xenotransplantation.( b+ H& i6 Y: @( j) M
Most multi-transgenic pigs have been produced by complex and time-consuming breeding programs using multiple
9 t9 t- q' A0 Ysingle-transgenic pigs. This study explored the feasibility of producing multi-transgenic pigs using the viral 2A peptide in, t! O, t. T ?) O% ]$ j9 }
the light of previous research indicating that it can be utilized for multi-gene transfer in gene therapy and somatic cell
/ p9 R' b. x% A! breprogramming. A 2A peptide-based double-promoter expression vector that mediated the expression of four fluorescent) C+ Z9 r! j5 t, J1 A- ]9 A7 s; |* W# N
proteins was constructed and transfected into primary porcine fetal fibroblasts. Cell colonies (54.3%) formed under G418) b2 [* {; S2 t/ q
selection co-expressed the four fluorescent proteins at uniformly high levels. The reconstructed embryos, which were n; B/ O! X* o' s; K
obtained by somatic cell nuclear transfer and confirmed to express the four fluorescent proteins evenly, were transplanted
, E3 Q9 q! ~% T; _4 J5 V/ Binto seven recipient gilts. Eleven piglets were delivered by two gilts, and seven of them co-expressed the four fluorescent6 k4 t5 X9 t# F+ M, S; Q* d% T
proteins at equivalently high levels in various tissues. The fluorescence intensities were directly observed at the nose, hoof
+ c9 v9 _' |9 m9 r0 Iand tongue using goggles. The results suggest that the strategy of combining the 2A peptide and double promoters |1 R7 V& p* N: w( f0 l
efficiently mediates the co-expression of the four fluorescent proteins in pigs and is hence a promising methodology to2 P7 V- G1 s2 J( } a
generate 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
