显微注射法制备转基因小鼠的技术研究

转基因技术是发生工程学或胚胎操作技术中的一部分。这一技术包括有受精卵的采集，受精卵的体外培养，胚胎移植等技术，转基因动物制作是其应用技术之一，它是将外源ＤＮＡ片段用显微注射法直接注入受精卵原核，使外源ＤＮＡ随机整合到寄主基因组中而形成转基因小鼠。我们建立完善这一技术的目的是在实验动物的生产中为转基因动物的生产提供稳定可靠的保证。我们将ｐＣＸＴＧＡＰ、ｐＣＳＬＮ等基因分别注入受精卵原核制备相应的转基因动物模型，并从中总结较佳的操作技术。结果经检测阳性转基因小鼠占出生幼鼠的１６３８％。我们认为不仅要有精细的微注射技术、动物的选用和饲育、严密的胚胎操作都是转基因动物制作成功的重要条件。

Study on Techniques of Making Transgenic Mice by Microinjection Method

ransgenic technique is one part of the development engineering or embryonic manipulation. This techniques include collecting embryos, in vitro culture of embryos and transferring embryos as a basic technology. Production of transgenic mice is one kind of application technology. The foreign DNA ware microinjected into nucle of fertilized one cell eggs and integrated into apparently random sit in the host genome. We produce transgenic mice by microinjection with NchW (Human prion protein gene),pCSLN(Sialidase-L gene),prbo(Bovine prion protein gene),CXTGAP(Human GM 2 activation protein gene),Prsh (Sheep prion protein gene) DNA fragments into the pronucleus of fertilized eggs. The DNA integrates on chromosomes at random and we got the positive result from the pups by PCR-Southern method. The average positive rate of transgenic mice is 16.38%. The present work proved that the main requirement of succeed on production transgenic mice should include precise technical skill, animal chosen and husbandry,manipulation