【摘要】 已知黑色素皮质素受体1(melanocortin-1 receptor,MC1R)是调控哺乳动物毛色的主要基因之一。羊驼是一种能生产纤维的家畜,有22种天然毛色。为了研究是否可以将羊驼的毛色基因应用到绵羊中,本文克隆了羊驼MC1R,发现羊驼MC1R与绵羊的相似性为88. 05%。将表达羊驼MC1R的质粒转染到绵羊黑色素细胞中,MC1R、小眼畸形相关转录因子(microphthalmia-associated transcription factor,MITF)、酪氨酸酶(tyrosinase,TYR)、酪氨酸酶相关蛋白2 (tyrosinase-related protein 2,TYRP2)在转录水平的表达量与NC组相比具有显著上升趋势,分别是对照组的3. 193±0. 063倍、2. 021±0. 129倍、3. 218±0. 056倍、3. 140±0. 108倍(P<0. 01)。Western印迹检测MITF、TYR、TYRP2在蛋白质水平的表达分别是对照组的1. 852±0. 117倍、3. 537±0. 13倍、2. 045±0. 163倍,具有显著差异(P<0. 01)。此外,绵羊黑色素细胞中羊驼MC1R的表达造成黑色素合成显著增加,是NC组的1. 715±0. 014倍(P<0. 01)。本研究结果表明,羊驼MC1R可用于增加绵羊黑色素生成过程中某些毛色基因的表达,为羊驼提供了可能的应用策略。更多还原

【Abstract】 Melanocortin-1 receptor( MC1 R) is known to be one of the main genes to control coat color in mammals. Alpaca,fiber-producing domestic animal,has 22 kinds of natural hair colors. To investigate whether hair color gene of alpaca can be applied to sheep,MC1 R from Alpaca( Alp-MC1 R) was cloned and results showed that the similarity between Alp-MC1 R and sheep was 88. 05% similarity. The plasmid expressing Alp-MC1 R was transfected into sheep melanocytes and a significant increase was found in mRNA expression of MC1 R,microphthalmia-associated transcription factor( MITF), tyrosinase( TYR) and tyrosinase-related protein 2( TYRP2) vs NC,which were 3. 193 ± 0. 063,2. 021 ± 0. 129,3. 218 ± 0. 056,3. 140 ± 0. 108 folds of the NC group respectively( P<0. 01). Western blot analysis showed that the protein expression of MITF,TYR and TYRP2 in Alp-MC1 R group were 1. 852 ± 0. 117,3. 537 ± 0. 13 and 2. 045 ± 0. 163 folds vs that in NC group respectively,with significant difference( P<0. 01). In addition,Alp-MC1 R expression in sheep melanocytes caused a significant increase in melanin synthesis,which was 1. 715 ± 0. 014 folds higher than that in the NC group( P<0. 01). Our results suggested that Alp-MC1 R could be used to increase the expression of some hair color genes in sheep during melanogenesis and provided a potential application strategy for alpaca.更多还原