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四种颜色的荧光蛋白以及合并后的蛋白 ; H p; H" `4 A5 F5 |! q' W9 t0 P& j% u% I9 p
8楼原文 感谢zyyzyy1985 提供作者: xuexi 时间: 2011-5-25 13:14
Abstract 9 M2 M7 m# N( J! j( {Multiple genetic modifications in pigs can essentially benefit research on agriculture, human disease and xenotransplantation.( A- ^1 F, F2 t7 `
Most multi-transgenic pigs have been produced by complex and time-consuming breeding programs using multiple y0 T% `$ w+ J: F4 O
single-transgenic pigs. This study explored the feasibility of producing multi-transgenic pigs using the viral 2A peptide in3 N% m" N- b0 V5 y' H3 j% m) W
the light of previous research indicating that it can be utilized for multi-gene transfer in gene therapy and somatic cell # q% h+ ]0 a# U- B9 w. }" Lreprogramming. A 2A peptide-based double-promoter expression vector that mediated the expression of four fluorescent - I. I l5 V6 p0 l3 u" j* e3 g% V) ]proteins was constructed and transfected into primary porcine fetal fibroblasts. Cell colonies (54.3%) formed under G418 $ c% z- r; X, V" O- W+ n5 Bselection co-expressed the four fluorescent proteins at uniformly high levels. The reconstructed embryos, which were$ ?. n; \0 q3 D9 G; R! h8 T
obtained by somatic cell nuclear transfer and confirmed to express the four fluorescent proteins evenly, were transplanted / T! Q* N' @$ l \5 W+ @' Ginto seven recipient gilts. Eleven piglets were delivered by two gilts, and seven of them co-expressed the four fluorescent/ _2 W% j# x( x% \9 h
proteins at equivalently high levels in various tissues. The fluorescence intensities were directly observed at the nose, hoof$ f, H, u: A `! K) ]
and tongue using goggles. The results suggest that the strategy of combining the 2A peptide and double promoters 3 x9 L0 G7 |9 D8 i) hefficiently mediates the co-expression of the four fluorescent proteins in pigs and is hence a promising methodology to ; w+ I. k1 [" R/ Tgenerate multi-transgenic pigs by a single nuclear transfer.作者: xiaomingxu 时间: 2011-5-28 11:36
恭喜赖老师实验室！作者: hormone008 时间: 2011-5-30 12:23
猪，或与之类似的大动物的转基因，一直是一个难攻克的技术难题！7 ]. _# e9 j1 a9 f1 b% d
而作为一个大动物的试验应用价值更是无法取代的！# D! G1 y' ?0 \/ f$ U" f% T% C
在全美国，也只有三家公司（实验室）能够做转基因猪。这是我的领导告诉我的。（她美国八年，刚刚回国，MD aderson）作者: hormone008 时间: 2011-5-30 12:24
