Abstract：
1. This study focused on the relationship between MITF mRNA expression and plumage colour in quail and the effect of promoter methylation on the expression of MITF mRNA.2. The CDS region of MITF mRNA was cloned by RT-PCR, followed by DNA sequencing. The RT-qPCR method was used to analyse the expression levels of MITF mRNA in dorsal skin tissue in Korean quail and Beijing white quail. The promoter region of the MITF gene was cloned, and the CpG island was predicted by the CpGplot program. The methylation levels of the CpG island were analysed using BS-PCR technology.3. Quail MITF mRNA contains a 1,476 bp complete ORF, which encodes a 492 amino acid residue protein. The MITF protein has no signal peptide or transmembrane region. The expression of MITF mRNA in dorsal tissue of Korean quail was significantly higher than that in Beijing white quail (p < 0.01). Abundant cis-elements and a 346 bp CpG island were found in the promoter region of the MITF gene. The average methylation level of the CpG island was 22 (22%) in Korean quail, and 46 (30%) in Beijing white quail (p < 0.05).4. The hypermethylation of the MITF gene promoter region in Beijing white quail resulted in a decrease in expression level, which was related to white feather colour.
摘要：
1.本研究主要探讨鹌鹑MITFmRNA表达与羽色的关系及启动子甲基化对MITFmRNA表达的影响。通过RT-PCR克隆MITFmRNA的CDS区，然后进行DNA测序。采用RT-qPCR方法分析韩国鹌鹑和北京白鹌鹑背部皮肤组织中MITFmRNA的表达水平。克隆了MITF基因的启动子区，CpG岛是由CpGplot程序预测的。使用BS-PCR技术分析CpG岛的甲基化水平。鹌鹑MITFmRNA含有1,476bp的完整ORF，它编码一个492个氨基酸残基的蛋白质。MITF蛋白没有信号肽或跨膜区。朝鲜族鹌鹑背部组织中MITFmRNA的表达明显高于北京白鹌鹑(p<0.01)。在MITF基因的启动子区域中发现了丰富的顺式元件和346bp的CpG岛。韩国鹌鹑CpG岛的平均甲基化水平为22（22%），北京白鹌鹑46只(30%)(p<0.05)。北京白鹌鹑MITF基因启动子区的高甲基化导致表达水平下降,这与白色羽毛的颜色有关。
