Kelley Clair M, Yergeau Donald A, Zhu Haiqing, Kuliyev Emin, Mead Paul E
Department of Pathology, St. Jude Children's Research Hospital, Memphis, TN, USA.
Methods Mol Biol. 2012;917:231-43. doi: 10.1007/978-1-61779-992-1_14.
The generation of transgenic animals is an essential tool for many genetic strategies. DNA "cut-and-paste" transposon systems can be used to efficiently modify the Xenopus genome. The DNA transposon substrate, harbored on a circularized plasmid, is co-injected into fertilized Xenopus embryos at the one-cell stage together with mRNA encoding the cognate transposase enzyme. The cellular machinery rapidly translates the exogenous mRNA to produce active transposase enzyme that catalyzes excision of the transposon substrate from the plasmid and stable integration into the genomic DNA.
转基因动物的产生是许多遗传策略的重要工具。DNA“剪切粘贴”转座子系统可用于有效修饰非洲爪蟾基因组。携带在环状质粒上的DNA转座子底物在单细胞阶段与编码同源转座酶的mRNA一起共注射到受精的非洲爪蟾胚胎中。细胞机制迅速翻译外源mRNA以产生活性转座酶,该酶催化转座子底物从质粒上切除并稳定整合到基因组DNA中。