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TARGETING GENE EXPRESSION TO THE WOOL FOLLICLE IN TRANSGENIC SHEEP

机译:将基因表达靶向到转基因绵羊的羊毛中

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摘要

To establish the feasibility of overexpressing foreign genes in the wool follicle, transgenic sheep were produced by pronuclear microinjection of a DNA construct consisting of a mouse ultrahigh-sulfur keratin promoter linked to the bacterial chloramphenicol acetyl transferase (CAT) gene. Four of 31 lambs born were transgenic. The overall efficiency of transgenesis was 1.1% of zygotes injected and transferred. Two transgenic rams were mated to nontransgenic ewes, and both transmitted the gene to their offspring in Mendelian fashion, CAT expression was found in the skin of one G0 ram and in 9 out of 26 transgenic G1 progeny, Two G1 lambs were sacrificed to study tissue specificity. Both had high levels of expression in skin but One had high expression in spleen and kidney,vith lower levels of expression in lung; the other had low expression in spleen, lung, and muscle. In situ hybridization demonstrated that transgene expression in the skin was confined to the keratogenous zone of the wool follicle cortex, Expression of CAT activity in skin was correlated with diet-induced or seasonal changes in the rate of wool growth. This keratin promoter appears useful for overexpressing factors in the wool follicle that might influence wool production or properties.
机译:为了建立在毛囊中过度表达外源基因的可行性,通过前核显微注射DNA结构构建了转基因绵羊,该DNA结构由与细菌氯霉素乙酰转移酶(CAT)基因连接的小鼠超高硫角蛋白启动子组成。出生的31只羔羊中有4只是转基因的。转基因的总效率为注入和转移的合子的1.1%。将两只转基因公羊与非转基因母羊交配,并以孟德尔方式将基因传递给它们的后代,在一只G0公羊的皮肤中发现CAT表达,在26个转基因G1后代中有9只在公羊中被牺牲以研究组织特异性。两者均在皮肤中高表达,但之一在脾脏和肾脏中高表达,而在肺中较低。另一个在脾脏,肺和肌肉中低表达。原位杂交表明,皮肤中的转基因表达被限制在毛囊皮层的角质形成区域。CAT活性在皮肤中的表达与饮食诱导的或季节性的羊毛生长速率相关。该角蛋白促进剂似乎可用于过表达毛囊中可能影响羊毛生产或特性的因子。

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