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MC1R和ASIP真核表达载体的构建及其对日本鹌鹑羽色的调控()。

Construction of MC1R and ASIP Eukaryotic Expression Vector and its Regulation of Plumage Color in Japanese Quail ().

作者信息

Li Yi-Xin, Zhang Xiao-Hui, Pang You-Zhi, Qi Yan-Xia, Zhao Shu-Juan

机构信息

College of Animal Science, Henan University of Science and Technology, P. R. China.

出版信息

J Poult Sci. 2019 Apr 25;56(2):84-90. doi: 10.2141/jpsa.0180058.

Abstract

The Japanese quail expresses polymorphism in plumage colors, including black, yellow, white, wild-type (maroon), and various intermediate colors through hybridization of quail with different plumage colors. The expression levels of MC1R and ASIP play important roles in the regulation of plumage colors in birds. In this study, the eukaryotic expression vector of pcDNA 3.1 + was used to analyze the effects of forced expression of MC1R and ASIP on the plumage colors of Japanese quail embryos. The constructed eukaryotic expression vectors of pcDNA 3.1 (+)-MC1R and pcDNA 3.1(+)-ASIP were transfected into wild-type Japanese quail embryos by Lipofectamine™ 2000 liposome at 6 days of incubation. After 3 days, the embryos were collected to analyze the plumage colors and the expression levels of , and genes in skin tissue. Forced expression of the gene by transfection of the pcDNA 3.1(+)-MC1R vector led to hyperpigmentation (similar to black plumage), whereas forced expression of the gene by transfection of the pcDNA 3.1(+)-ASIP vector led to hypopigmentation (similar to white plumage) in wild-type quail embryos. Two kinds of ASIP alternative splicing (ASIP1 and ASIP2) were found in Japanese quail, which did not have a significant effect on the plumage color or the main motifs of the ASIP protein. This study indicated that the black plumage color may be caused by increased production of MC1R and the white plumage color may be caused by increased production of ASIP in Japanese quail.

摘要

日本鹌鹑的羽毛颜色存在多态性,包括黑色、黄色、白色、野生型(栗色),以及通过不同羽色鹌鹑杂交产生的各种中间颜色。MC1R和ASIP的表达水平在鸟类羽毛颜色的调控中起着重要作用。在本研究中,使用pcDNA 3.1 +真核表达载体来分析MC1R和ASIP的强制表达对日本鹌鹑胚胎羽毛颜色的影响。构建的pcDNA 3.1(+)-MC1R和pcDNA 3.1(+)-ASIP真核表达载体在孵化第6天时通过Lipofectamine™ 2000脂质体转染到野生型日本鹌鹑胚胎中。3天后,收集胚胎以分析羽毛颜色以及皮肤组织中相关基因的表达水平。通过转染pcDNA 3.1(+)-MC1R载体强制表达该基因导致色素沉着过度(类似于黑色羽毛),而通过转染pcDNA 3.1(+)-ASIP载体强制表达该基因导致野生型鹌鹑胚胎色素沉着不足(类似于白色羽毛)。在日本鹌鹑中发现了两种ASIP可变剪接(ASIP1和ASIP2),它们对羽毛颜色或ASIP蛋白的主要基序没有显著影响。本研究表明,日本鹌鹑中黑色羽毛颜色可能是由于MC1R产量增加所致,白色羽毛颜色可能是由于ASIP产量增加所致。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/99a9/7005409/ae1c715bc8b1/jpsa-56-84-g001.jpg

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