Study On The Exon 1 Splicing And Promoter Activity Area Of Goat Agouti Gene

The study designing primers with the Agouti gene sequence of Yunnan black goat w hich was a template, using black and white goat genomic DNA as a sample of the Agouti gene partial sequences were amplified and obtained the Agouti gene partial sequences.On the basis of the skin of different hair color goat of agouti gene 5 ’untranslated regi on significantly different types of exon 1 spliceosome, I designed primers.The splicing exp erience of the black and white colors goat were adopted.. The results showed that: the spli cing type of the selected individual goats were It It’.The homology of two parts of the Goat genomic sequence(Accession number: AJPT01136617.1) and two predict promoter area sequences of cattle were high, therefore design ed primers to amplify target fragment A and B, while builded the two luciferase reporter g ene plasmid PGL3-basic-A and PGL3-basic-B. And the construction of luciferase reporter gene plasmid was transiently transfected into human skin melanoma cell line A375(skin) cells and 293 T cells(non skin cells) to detect the two objective fragments of Agouti gene promoter activity. The results were analyzed with SPSS software. The results showed that the fragment of the constructed plasmid and negative controls than does not exist significa nt differences, indicating that the obtained two fragments do not have promoter activity.