Clone Of Bos D2-like Gene In Sheep And Its Effects On Coat Color Related Gene Expression

The target gene28954, which was obtained by transcriptome sequencing, was a potential new gene which was related to black and white coat color in ovis aries. First, the research proved the expression difference of28954gene between the ovis aries skins with black and white coat color by real-time fluorescent quantitative PCR. The result showed that the expression of28954mRNA in the black ovis aries skin was1.9185times of that in white ovis aries. It suggested that the28954gene had a possible correlation with coat color.To further understand the28954gene, we did a bioinformatics analysis for its cDNA sequence. In the beginning, the named homologous gene wasn’t found. But with the updating of NCBI database, we found the28954gene was one of transcripts of Bos d2-like （LOC101116799）. However, till now, we did not find any research reports about Bos d2-like （including the researches related to coat color）.To further study the relationship between Bos d2-like and ovis coat color, we designed a pair of primers of Bos d2-like（LOC101116799） according to the ovis aries genomic sequence （NW₀04080190） which was logged in NCBI GenBank., the557bp coding region sequence of ovis aries Bos d2-like was amplified for the first time. Then, the amplified sequence was cloned to pcDNA3.1eukaryotic expression vector, called pcDNA3.1-Bd21. It was transfected into melanocytes, and detected the expression of TYR, TYRP1, MC1R, MITF mRNA were detected by QRT-PCR, in order to reveal the function and molecular mechamism of Bos d2-like in melanogenesis. The QRT-PCR showed that the overexpression of Bos d2-like mRNA was1.923times compared with the control group （P<0.05）.The expression of MCI R mRNA in Bos d2-like overexpression group decreased0.394times of that in the control group （P<0.05）, while the expression of TYR, TYRP1and MITF m RNA in Bos d2-like overexpression group tended to decline but had no significant differences, compared with the control group. The results suggested that Bos d2-like（LOC101116799） could downregulate the transcriptional level of MC1R in melanocyte and then might affect the biosynthesis of melanin in melanocyte.