Cloning And Expression Of BMP15and BMP2B Genes In Cyprinus Carpio VAR. Jian

Cyprinus carpio var. jian, as China’s most important aquaculture specie of common carp, was favored by lots of consumers and farmers duing to its better meat quality and taste, higher feed transformation efficiency, docile temperament soft characteristic, stronger adaptability and disease resistance, easier to domesticate and catch, breeding reproduced and cultivating babies by itself and its higher stable genetic stablility characteristic,and so on. But the thorn between intrermusclar bones has had negative influence on its deep production processing and its economic value. BMP genes are were involved in bone formation, playing an important role in repairing bone defect and promoting the healing. However, whether BMPs had a important role in formation of fish intermuscular bone, occurrence mechanism, distribution action was still unclear. Hence, this experiment was conducted to research the relationship between BMP15, BMP2B struction, impression and e within intermuscular bon and without in fish in different species through use RT-PCR and Real-time PCRThe results as follows:1The full length cDNA sequence of BMP15gene and the partial sequence of BMP2B gene, encoding359and266amino acids respectively were cloned. Amino acid sequences have higher homology with other species.2The analysis of amino acids of BMP15gene showned that TGF-β domain and maximum hydrophobicity2.711were got; protein structure analysis showned that a transmembrane domain and a protein signal peptide were predicted; secondary structure prediction illustrated that coil has the largest share of65.74%, Alpha-Helix is21.70%, and the Beta-Strand were13.09%at least; Besides these, BMP15gene may be located mitochondria mostly, which had14Ser,6Thr,3Tyr and may be regarded as phosphorylated sites of protein kinase has and a N-glycosylation sites, and not O-glycosylation sites.The BMP2B gene amino acids sequence analysis was also done, the result suggested that it had two lower complexity structure of domains, a TGF-β domain and maximum hydrophobicity1.689; Compared to BMP15, BMP2B had a transmembrane domain, protein signal peptide and coil had the largest share of60.90%, while Beta-Strand was30.45%,and the Alpha-Helix was8.65%; BMP2B was located cytoplasm which was different from BMP15and had7Ser,0Thr,3Tyr, phosphorylated sites of protein kinase possibly; similarly, a N-Glycosylation sites and not O-glycosylationl sites were also predicted.3Phylogenetic tree was built, in which Jian carp BMP15was clustered with zebrafish BMP15firstly, and then clustered with Allogynogenetic Silver Cruican Carp, Europe the sea bass BMP15, and mammals BMP15also cluster obviously, and then get together,on comparison shows the Jian carp BMP15cluster with several cyprinid fishs BMP15at first,phylogenetic relationship is near.The Jian carp BMP2B cluster with belong to the carp families of zebrafish BMP2B at first, and demonstrate that the phylogenetic relationship with zebrafish is the nearest. Each species of fish BMP2B cluster, and mammals, BMP2B also obvious cluster, the y are divided into two branches obviously.4The Real-timePCR was used to analysis the expression profiles of BMP15and BMP2B in different common carp tissues.uncovered that:BMP15genes are expressed in eight organization, but expression level is not the same. It expressed the highest in ovarian, liver and muscle expressed in second one, once again in bowel, brain and gills while heart and spleen expressed in the lowest. BMP2B genes are expressed in eight organization tissues, but expression level is not the same. It expressed the highest in muscle tissue; ovarian and liver expressed in second one, once again in spleen, gills and bowel, while heart and brain expressed in the lowest.In this paper, the Jian carp BMP15and BMP2B genes were cloned successfully using the RT-PCR and RACE technology, and the expression profiles analysis of such two genes were also observed using Real time PCR. These studies may contribute to the further research BMP15and BMP2B gene function, regulation, and also laid the foundation for further study between the formation of fish intermuscular bone happened mechanisms.