Cloning And Mapping Of By Fluorescence In Situ Hybridization(FISH) Of BMP-2 Gene From Inner Mongolia Cashmere Goat

In this study,the genome DNA were extracted from blood in Cashmere goat.The sequence were obtained by PCR and cloned into pGM18-T vector .The positive clones were sequenced.Then we compared the sequences of BMP-2 gene fragments among goats,sheep and other animals that have been submitted to GeneBank. The method of bioinformatics was used to aligment of both their nucleotide and amino acid sequence of Cashmere goat. Furthermore we made the probe with DIG using the cloning product of Cashmere goat, for studying the map of BMP-2 in Cashmere of Cashmere goat by fluorescence In situ hybrition. The results would enrich gene map and then offered the theoretical support to understand the effect of gene message of BMP-2 on breeding of Cashmere goat.The results showed as following:1. Obtained the sequence of the exon of bmp2 which spaned 1751bp. The phylogenetic trees among goat, sheep, cow, pig, human and macaca conformted to taxonomy of tradition. It is suggested that BMP-2 gene can represent the character of Cashmere goat.2. The replacement of Exon 3 of BMP-2 among goat, sheep and cow was only nucleotide. It was found 4 mutant among goat and sheep,99.6% identity between cashmere goat and sheep. It was found 13 mutant among goat and cow,98.1% identity between cashmere goat and cow. The sequence of the BMP-2 gene's Exon 3 was translated to amino acid sequence. The results of aligment of the amino acid showed that it was found 5 mutant among Cashmere goat, sheep and cow, but only 1 mutant between goat and sheep, the identity of goat and sheep was very high. It is thus clear that, the replacement of BMP-2 gene between goat and sheep is samesense mutation, furthermore explanation of BMP-2 gene'conservatism is very better.3. The recombinant plasmid were marked, accroding to the guider of kit, then made a probe. BMP-2 gene was maped into chromosome 13q15 of Cashmere goat by fluorescence In situ hybridization.