Bai Ding-Ping, Yang Ming-Ming, Chen Yu-Lin
College of Animal Science and Technology, Northwest Agriculture and Forest University, Shaanxi, PR China.
Biosci Biotechnol Biochem. 2012;76(5):933-7. doi: 10.1271/bbb.110939. Epub 2012 May 7.
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.
最近发现PiggyBac（PB）转座子在多种生物体中具有功能。为了验证并开发其在绒山羊中的应用，构建了一个包含供体质粒和辅助质粒的PB转座子系统，其中供体质粒中的增强型绿色荧光蛋白（EGFP）基因用作报告基因。用该PB转座子系统转染绒山羊胎儿成纤维细胞（GFFs），并测定基因转移效率。与随机整合相比，PB介导的EGFP在GFFs中的表达水平提高了7.78倍，证实成功构建的PB转座子系统介导了外源基因在GFFs中的高效整合。为了进一步研究PB介导的整合情况的特征，检测了PB在山羊基因组中的整合位点分布。结果表明，PB倾向于整合到山羊基因组中富含AT的区域。因此，本研究证实了PB转座子在GFFs中的功能，并为生产转基因山羊提供了一种潜在的遗传工具。
