Freitas V J F, Serova I A, Andreeva L E, Júnior E S Lopes, Teixeira D I A, Cordeiro M F, Rondina D, Paula N R O, Arruda I J, Verde J B Lima, Dvoriantchikov G, Serov O
Laboratory of Physiology and Control of Reproduction, State University of Ceará, Av. Paranjana, 1700, 60740-000 Fortaleza, CE, Brazil.
Genet Mol Res. 2003 Jun 30;2(2):200-5.
This pilot project was designed to determine if normal kids could be produced after microinjection in pronuclear embryos and subsequent transfer to recipients in a transgenic goat program in Brazil. Twelve donors of the Saanen breed and 17 recipients of an undefined breed were used. The estrus of both donors and recipients was synchronized by a standard progestagen treatment and superovulation obtained by six pFSH injections. Donors in estrus were mated with fertile Saanen bucks. Zygotes were recovered surgically by flushing oviducts. The recovered zygotes with visible pronuclei were microinjected with 500 to 1000 copies of the human G-CSF gene. Two or four embryos were surgically transferred into the oviducts of recipients. One recipient became pregnant and two kids were born. No transgenic goat was identified after PCR analysis. Even though transgenic goats were not obtained, this experiment establishes the basis of a synchronization and superovulation regimen for use in goats raised in Brazil, for the purpose of collecting and manipulating the pronuclear embryos. This project also showed that microinjected one-cell goat embryos can survive to produce live young following surgical transfer.
这个试点项目旨在确定在巴西的转基因山羊项目中,对原核胚胎进行显微注射并随后移植到受体体内后,是否能够培育出正常的幼崽。使用了12只萨能品种的供体和17只品种不明的受体。供体和受体的发情期通过标准的孕激素处理进行同步,通过六次注射促卵泡素(pFSH)实现超数排卵。处于发情期的供体与可育的萨能公羊交配。通过冲洗输卵管手术回收受精卵。将带有可见原核的回收受精卵显微注射500至1000份人类G-CSF基因。通过手术将两个或四个胚胎移植到受体的输卵管中。一只受体怀孕并产下两只幼崽。经聚合酶链反应(PCR)分析后未鉴定出转基因山羊。尽管没有获得转基因山羊,但该实验确立了在巴西饲养的山羊中用于同步发情和超数排卵方案的基础,以便收集和操作原核胚胎。该项目还表明,显微注射的单细胞山羊胚胎在手术移植后能够存活并产下活体幼崽。