整合人mAAT基因转基因山羊的获得

Production of Transgenic Lamb Integrated with Modified Human Anti-trypsin Gene

旨在通过动物转基因技术开发奶山羊的乳腺生产治疗人肺纤维性囊肿的新药──人抗 胰蛋白酶因子（人ｍＡＡＴ）。选择繁殖机能优良的奶山羊５６只作为供体，采用ＦＳＨ＋ＬＨ做超 数排卵，获取原核期胚胎，并进行显微注射和鲜胚移植．３月和５月两个月份超排处理后超排 效果明显好于１２月份，分别可获得平均排卵１９．５０、２１．７０和１６．０６枚，受精卵数分别为４．３１、 ６．４８和３．５７枚．显微注射后胚胎移植给供体或自然发情的受体，移植受胎率分别为１８．１８％和 ２５．００％．所生羔羊２９只，对转入外源基因编码区Ｎ－末端和３’调控区部分序列进行ＰＣＲ、 ＰＣＲ－Ｓｏｕｔｈｅｒｎ和Ｓｏｕｔｈｅｒｎ检测，检出阳性转基因羊羔４只，基因的整合效率为１３．７９％．

This experiment is to produce the human mAAT(modified anti-trypsin) which cures the emphysema specifically through mammalian galactophore of transgenic goat.56 goats were selected as donor for superovulation by FSF+ LH microinjection in this experiment.The pronucleic embryos were injected with human mAAT gene after fertilization in vivo, and transferred to the donors of receptors directly. The superovulation was better in March and May than in December with the number of ovulation of 19.50, 21.70 and 16.06, and number of fertilized embryos of 4.31, 6.48 and 3.57 peranimal peranimal . The pregnant rates were 18.18% and 25% respectively after transferred to donors and receptors with natural estrus. The donors also can be used as the embryo receptor with no remarkable decrease of Pregnant rate.29 lamb were labored. 4 positive transgenic lamb were checked by PCR, PCR-Southern and Southern analysis. The integrated efficiency of foreign DNA was 13.79% with microinjection of high copy number of foreign DNA fragment.