PiggyBac transposon-mediated gene transfer in Cashmere goat fetal fibroblast cells.
PiggyBac (PB) has recently been found to be functional in various organisms. To verify and exploit its application in the cashmere goat, a PB transposon system including donor and helper vector of was developed, in which the EGFP gene in donor of vector was used as reporter. Cashmere goat fetal fibroblasts cells (GFFs) were transfected with the PB transposon system and the efficiency of gene transfer was determined. Compared with random integration, PB-mediated EGFP expression levels increased 7.78-fold in the GFFs, confirming that the PB transposon system constructed successfully mediated efficient foreign gene integration in the GFFs. To further investigate the characteristics of PB-mediated integration instance, PB integration site distribution in the goat genome was examined. The results showed that PB had a preference for AT rich regions of the goat genome. Thus this study confirms the function of PB transposon in GFFs and provides a potential genetic tool for producing transgenic goats.
College of Animal Science and Technology, Northwest Agriculture and Forest University, Shaanxi, PR China.
Ding-Ping
DP
Bai
Ming-Ming
MM
Yang
Yu-Lin
YL
Chen
D020856:AT Rich Sequence; D000818:Animals; D004251:DNA Transposable Elements; D005333:Fetus; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D016678:Genome; D006041:Goats / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D010957:Plasmids; D014162:Transfection
D004251:DNA Transposable Elements; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins