世界首例可发红黄绿青四种荧光的克隆猪（附原文）

xuexi

Abstract
  I, j: u/ U) y" [5 x9 lMultiple genetic modifications in pigs can essentially benefit research on agriculture, human disease and xenotransplantation.
5 B# t/ j/ _, bMost multi-transgenic pigs have been produced by complex and time-consuming breeding programs using multiple/ S. I& _5 ^" m* q1 j) i
single-transgenic pigs. This study explored the feasibility of producing multi-transgenic pigs using the viral 2A peptide in6 a# u( ?( ~. I2 Y' k  L
the light of previous research indicating that it can be utilized for multi-gene transfer in gene therapy and somatic cell
5 J# U$ \- `+ t2 n! preprogramming. A 2A peptide-based double-promoter expression vector that mediated the expression of four fluorescent
" a* r$ }/ O0 B+ J- f  }proteins was constructed and transfected into primary porcine fetal fibroblasts. Cell colonies (54.3%) formed under G4189 \4 h8 U3 k8 \% |/ J( d
selection co-expressed the four fluorescent proteins at uniformly high levels. The reconstructed embryos, which were, x0 Y0 P2 n7 U9 c! A5 {! z
obtained by somatic cell nuclear transfer and confirmed to express the four fluorescent proteins evenly, were transplanted/ U0 r. a7 [# F9 x* ^! l' G8 m
into seven recipient gilts. Eleven piglets were delivered by two gilts, and seven of them co-expressed the four fluorescent$ j: n* S- D% |7 Z; S' ?" L+ y) H) p
proteins at equivalently high levels in various tissues. The fluorescence intensities were directly observed at the nose, hoof4 x8 p8 q  P% z; B
and tongue using goggles. The results suggest that the strategy of combining the 2A peptide and double promoters
* X6 [6 \; ]& ~2 n1 \3 `, M4 C3 s& Jefficiently mediates the co-expression of the four fluorescent proteins in pigs and is hence a promising methodology to
1 t. }  q$ b' G( c" k# r4 m* kgenerate multi-transgenic pigs by a single nuclear transfer.