睾丸注射法制备携带徐淮山羊过氧化氢酶体激活增殖受体γ基因转基因羊的研究  

Study on Generation of Xuhuai Goat PPARγ Gene Overexpression Transgenic Sheep by Testicular Injection

作　　者：朱才业[1] 李伟[1] 朱睿[1] 韦光辉[1] 邱峰龙[1] 秦玉蓉 张亚妮[1] 李碧春[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009 [2]南京金斯瑞生物科技有限公司,江苏南京210000

出　　处：《中国畜牧兽医》2013年第6期153-157,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家转基因重大专项(2008ZX08008-003;2009ZX08008-003B)

摘　　要：为探究睾丸注射法制备转基因动物的可能性,将携带有徐淮山羊过氧化氢酶体激活增殖受体γ(PPARγ)和绿色荧光蛋白标签的重组载体经脂质体包裹后,将线性化的重组质粒pEGFP-PPARγ通过随机打点注射入性成熟的湖羊睾丸内,采用常规PCR、基因组DNA点杂交及蛋白免疫印迹技术检测外源PPARγ基因在湖羊体内稳定整合的能力,并检测F1代阳性个体屠宰性能、肌内脂肪含量、器官重量及肉品质。F1代羔羊检测转基因羊阳性率为13.7%,证实外源基因在转基因后代体内成功表达。阳性个体胴体重和屠宰率极显著低于对照组个体(P<0.01),阳性个体骨肉比极显著高于对照组个体(P<0.01);阳性个体背最长肌脂肪含量显著高于对照组个体(P<0.05),腰肌内脂肪含量与股二头肌内脂肪含量差异不显著(P>0.05);阳性个体肺脏和小肠极显著大于对照组个体(P<0.01),阳性个体脾脏显著大于对照组个体(P<0.05),阳性个体心脏和肺脏显著小于对照组个体(P<0.05),肝脏、肾脏、胃、睾丸、皮、头之间差异不显著(P>0.05);阳性个体股二头肌嫩度显著低于对照组个体(P<0.05),pH、肉色、失水率之间差异不显著(P>0.05)。To explore the possibility of transgenic animals by testicular injection, the Xuhuai goat peroxisome proliferator- activated receptor expression vector pEGFP-PPARγ was using lipofection method, then the linearized recombinant plasmid pEGFP-PPAR7 by random dot injected into sexually mature Hu sheep testis. Conventional PCR, genomic DNA dot blotting and Western blotting detection the ability of exogenous PPARγ gene in Hu sheep stable integration. Detection positive individ- uals and negative individuals of F1 generation of performance of slaughter, the intramuscular fat content, organ weight and meat quality. Positive rate of transgenic sheep was to 13.7%, initially confirmed that the foreign gene PPAR7 could be inte- grated into the genome of Hu sheep and inherited to the next generation. Carcass weight and slaughter rate of positive individu- als were significantly lower （P〈0.01） than negative individuals in F1 Generation, The proportion of flesh of positive individu- als was significantly higher （P〈0.01） than negative individuals in F1 Generation~ the fat content of longissimus muscle of posi- tive individuals was higher （P〈0.05）than negative individuals, the fat content of psoas and biceps femoris were no significant difference （P〈0.05） ;positive individuals lung, small intestine were significantly higher （P〈0.01） than negative individuals, positive individuals spleen was higher （P〈0.05） than negative individuals, positive individuals heart, lung were lower （P（ 0.05） than negative individuals, liver, kidney, stomach, testis, skin, and head were no significant difference （P〈0.05）;posi- tive individuals was lower （P〈0.05） than negative individuals, no significant difference between the p H, meat color, water loss rate.

关 键 词：过氧化氢酶体激活增殖受体 睾丸注射 转基因羊 

分 类 号：S827[农业科学—畜牧学]