The Sequence Analysis And Expression Level Detection Of Skincolor Related Gene MC1R, MITF And Agouti In Yak

The coat colour of Yak affects the economic value of its wool. We choose Gannan yak and Tianzhu white yak for studying the genetic reasons in coat color formation, in order to understand the mechanisms. Cloning and sequencing yak MC1 R, MITF and Agouti gene by using molecular biology techniques. The MC1 R, MITF and Agouti gene related bioinformatics and polymorphisms were analyzed by related software. The expression levels of MC1 R, MITF and Agouti m RNA in different color yak skin tissues were examined by Quantitative PCR. The results were as follows:(1)The CDS of yak MC1 R, MITF and Agouti gene were cloned. The CDS of MC1 R mRNA is 1021 bp, which encoding 317 amino acids. MC1 R protein belongs to the hydrophilic protein. It is a hydrophilic protein, which has no signal peptide but have oglycosylation site and seven trans-membrane regions. The CDS of MITF mRNA is 1460 bp, which encoding 413 amino acids. MITF-M has a MLEMLEYNHY. It is a hydrophilic protein, which has no signal peptide and no trans-membrane regions. MITF-M contains 25 phosphorylation sites. The CDS of Agouti mRNA is 593 bp, which encoding 133 amino acids. It is a hydrophilic protein. It has no signal peptide and no transmenbrane regions. ASIP contains 5 phosphorylation sites.(2) The polymorphisms of MITF gene and Agouti gene CDS were identificated by DNA pools and direct sequencing. The gene sequencing showed that the yak MITF gene and Agouti gene was without any polymorphisms.(3) The semi-quantitative analysis of MC1 R gene and MITF-M showed that MC1 R gene and MITF-M subtypes only expression in the skin tissue. Quantitative PCR was employed to examine the expression levels of MC1 R, MITF and Agouti mRNA in different color yak skin tissues. Agouti gene was wildly expressed in ovary, muscle and skin tissues, and skin expression was the highest. The expression level of Agouti mRNA in skin tissues between dark yak and white yak were no significant difference. MC1 R gene mRNA in black skin was significant higher than that in white skin. It is implied that high mRNA expression of MC1 R gene is conducive to the deposition of melanin in the coat. Also, the expression level of MITF-M mRNA in skin tissues between dark yak and white yak were extremely remarkable. It is implied that high mRNA expression of MITF gene is conducive to the deposition of melanin in the coat.