Study On Differental Expression Of MLPH Gene And The Va Riations Of The Intron Mutation Among Different Colour G Oat Skin Tissue

According to published in Genbank goat MLPH gene sequence of CDS (EU332403)design primers for amplification, amplification length was104bp. From literature choosingof housekeeping gene GAPDH as internal amplification, gene amplification length was107bp. Using Real-time fluorescent quantitative PCR method, through Bio-Rad iQ5software to get the initial concentration, using relative quantitative2-ΔΔCtanalysis methodto calculate the MLPH gene in goats different coat color of skin tissue differentialexpression of mRNA: white (1.000)> black (0.204)> brown (0.094). Results showed, inthe white samples, the expression of MLPH gene was the highest, black was in the middleand brown was at least. Between white and the other two coat color difference significant(P <0.05).Measured by this test according to this research group published in Genbank(EU316218) part goat MLPH gene sequences, respectively in intron7part of the sequence,introns14and exons15partial sequence were designed two pairs of primers foramplification, amplification length is516bp and791bp respectively. In these twofragments amplified, multiple mutations were found. So we sequenced4breeds goatsamples and analysised120individuals for population genetics. Further analysis ofmutations and MLPH genetic relationship were studied.In part goat MLPH gene intron7pieces, six mutations were found, for respectively ing.G8067C, g.T8068C,g.C8104G,g.G8151A,g.A8352G,g.A8362G.Among g.G8067C andg.T8068C,g.A8352G and g.A8362G respectively showed the complete chain phenomenon.Analysis of Site g.G8067C and g.T8068C,we got two haploid type GT (65.8%) and CC(34.2%),from site g.A8352G and g.A8362G, we also got two haploid type AA (76.2%) andGG (23.7%). Site g.G8067C,g.T8068C,g.G8151A,g.A8352G,g.A8362G are related to colordilution gene MLPH goats. Six mutations of comprehensive analysis of Jining grey goatsand Liaoning down producing goat close relatives, and Leizhou native goat distantrelatives, the clustering result and location are conform to the results of the analysis.In goats MLPH gene intron14and exon15part pieces, seven mutations were found.Six mutations were on intron14, respectively g.G16669T, g.T16710A, g.C16711A,g.A16761G, g.C17041T, g.G17075C. The right to g.C16711A,g.A16761G and g.C17041T showed the full right to chain phenomenon, and two haploid type CAC (77.5%) and AGT(22.5%) were obtained; In exon15, only one mutation was found in a loci g.G17298T. Siteg.C16711A、g.A16761G and g.C17041T were associated with a goat color dilution. Theintrons14on the comprehensive analysis of the six mutations and intron7consistentresults. Goats different varieties by genetic loci and the influence of different geographicalenvironment interaction.