生殖生物学理论与相关技术：哺乳动物克隆技术.pptVIP

世界首例体细胞克隆兔及转基因克隆兔 2007克隆出世界首例转基因克隆兔 Transgenic Res 18(2):227-35,?2009 Apr (TRAGonline) doi:10.1007/s11248-008-9227-y 世界首例转基因克隆兔在沪诞生(图)2007-12-14 08:50　来源：解放日报 实验首先取兔耳组织培养获得所需要 的成纤维细胞，经过３～４次传代， 再将一种从特殊水母中提取的绿色荧光 蛋白基因转染到兔子成纤维细胞中， 再挑选出发绿色荧光的转基因体细胞， 将其细胞核移植到成熟的去核兔子 卵母细胞中，最终构成了转基因胚胎。 胚胎经过手术，移植入“代孕兔”体内。 经过３０～３１天孕期，通过给代孕 兔剖腹产获得幼兔，再经人工辅助哺育， 顺利断奶等一系列的过程，最终成功 获得了这只转基因克隆兔。转基因克隆 兔由于基因与原来兔子完全一样，加上 有发绿光的标记基因，便于鉴定，可以 尝试转入或转出某种基因，研究特定基 因和相关疾病的关系。 Glucose of the transgenic rabbit without Dox mmol/L 2h after food 2h before food 2h after food 2h before food I1-2-1 8.1 6.8 7.7 6.9 I2-1-1 10.5 5.2 9.9 4.9 I2-2-1 7 6.5 10.3 5.7 Wild1 7.8 6.8 6.8 5.8 Wild2 7.1 5.4 7 6.9 The transgenic rabbit I2-2-1 was injected Dox two times (0d,4d) .4d after the first injection，the glucose was 30mmol/L. Both prOG transgenic SCNT embryos and F1 prOG transgenic embryos derived from adult transgenic rabbit expressed green fluorescence at the morula and blastocyst stages. EGFP expression in the gonads of Oct4-EGFP transgenic fetuses EGFP expression in fetal tissues or organs at different developmental stages The fetuses at 15 dpc and 27 dpc were isolated.(A) green fluorescence was detected in several cells between the two genital ridges in the fetus at 15 dpc . (B) EGFP expression in the testis of a 27-dpc fetus. Right, testis from a prOG transgenic fetus; left, testis from a normal fetus. (C) EGFP expression in the ovary of a 27-dpc fetus. （Int. J. Dev. Biol 2013） Traditional HPRT knockout Vector transferred into adult rabbit fibroblast More than 80 fibroblast clones were selected after G418 treatment , but no one of them could survived in culture medium with 6-TG. 11月2日，医学院实验动物科学部陈学进研究小组的最新研究成果在Scientific Reports（IF=5.578）杂志上发表，研究论文题为“Generation of hypoxanthinephosphoribosyltransferase geneknockout rabbits by homologousrecombination and gene trappingthrough somatic cell nucleartransfer”（利用核移植及同源重组和基因捕获技术获得HPRT基因敲除兔）。 /articles/srep16023 2007年克隆出上海首例克隆猪 上海诞生首例克隆猪（图）2007年07月2