Expression And Localization Of Gas In Different Color Coats Of Sheep Ears

MC1R (melanocortin 1 receptor) was one of the important genes that involved in regulating different coat color of mammalian, MC1R was present in the surface of melanocytes, regulated the synthesis of melanin. MC1R was a G protein coupled receptor, which was activated by heterotrimeric G protein. Gas located in downstream of the heterotrimeric G protein, which had different expression in different coat color. It regulated pigment types of melanocytes by melanocortin signaling pathway. Thus, Gas may play an important role in the formation of coat color.To improve our understanding of Gas expression levels and pattern in the skin of black sheep compared with white sheep, we designed primers to amplify the target sequence of sheep Gas gene, analyzed the expression levels through quantitative real time PCR (qPCR),Western blot and immunohistochemistry. It provided the theoretical basis for the further study of the formation mechanism of the coat color about Gas.The following test results:1.As the sequence of Gay published by the Genebank of NCBI and based on the conserved regions of the sequence,we designed and synthetic the primer by primer design principles. Partial sequence of Gas was amplified by reverse transcription PCR (RT-PCR), The length of sequence was 139bp.In order to explore the correlation between the expression level and coat color, real-time quantitative PCR was applied for examining the mRNA expression level of Gas in sheep ear skin of different coat colors.he partial sequence of Gas gene was successfully amplified,The length of sequencing was 139bp. Results were shown as:The gene expression quantity of black skin of sheep ear was 2.7185±0.3565, The gene expression quantity of white skin of sheep ear was 1.2542±0.1132. The expression of black skin was significantly higher than that of white skin.It showed that there was a certain correlation between Gas and coat color.2.The Gas was demonstrated to express in outer root sheath and dermal papilla of hair follicular by immuohistochemistry.3.The average protein expression of Gas in black ear skin was significantly higher than that in white ear skin by Western blotting (P<0.05).In a word.there are differences in expression of Gas in sheep ear's black skin and white skin.it concludes that Gas gene is related to the formation of animal hair color.