Schnieke A E, Kind A J, Ritchie W A, Mycock K, Scott A R, Ritchie M, Wilmut I, Colman A, Campbell K H
PPL Therapeutics, Roslin, Midlothian, EH25 9PP, Scotland, UK.
Science. 1997 Dec 19;278(5346):2130-3. doi: 10.1126/science.278.5346.2130.
Ovine primary fetal fibroblasts were cotransfected with a neomycin resistance marker gene (neo) and a human coagulation factor IX genomic construct designed for expression of the encoded protein in sheep milk. Two cloned transfectants and a population of neomycin (G418)-resistant cells were used as donors for nuclear transfer to enucleated oocytes. Six transgenic lambs were liveborn: Three produced from cloned cells contained factor IX and neo transgenes, whereas three produced from the uncloned population contained the marker gene only. Somatic cells can therefore be subjected to genetic manipulation in vitro and produce viable animals by nuclear transfer. Production of transgenic sheep by nuclear transfer requires fewer than half the animals needed for pronuclear microinjection.
将新霉素抗性标记基因（neo）和设计用于在羊奶中表达编码蛋白的人凝血因子IX基因组构建体共转染绵羊原代胎儿成纤维细胞。两个克隆转染子和一群对新霉素（G418）有抗性的细胞用作供体，将其细胞核移植到去核卵母细胞中。六只转基因羔羊存活出生：三只由克隆细胞产生，含有因子IX和neo转基因，而三只由未克隆群体产生，仅含有标记基因。因此，体细胞可以在体外进行基因操作，并通过核移植产生存活的动物。通过核移植生产转基因绵羊所需的动物数量不到原核显微注射所需动物数量的一半。
