Study Of Liaoning Cashmere Goat Gene Biological Characteristics

Liaoning new breed of cashmere goat is a valuable genetic resource in china. And itsproducts，an important economic value,such as cashmere, meat,skin and hair.Thus,study onthe biological characteristics of Liaoning cashmere goat genes had great significance.Studies have found that melatonin can regulate the growth of cashmere goat hair.However, melatonin has not been reported in Liaoning new breed of cashmere goat geneexpression. Therefore, Liaoning new breed of cashmere goat was reacted as the researchobject for the first time, exploring the effects of melatonin decorin (DCN) GMP reductase2(GMPR2), and OCIAD2(ovarian cancer immune response to the antigen-antibody) three newgenes in Liaoning cashmere goats ear fibroblasts. The final concentration of melatonin was0.02g/L,0.2g/L,1g/L respectively in Liaoning new breed of cashmere goat ear fibroblasts,and the concentration treated cells were cultured for24h,48h,72h.At last quantitative PCRwas used to detect the expressin of the there genes and compare with Atin. The results show:(1) GMP reductase2in melatonin48,72hours, the concentration of0.2g/L, the mostsignificant effect (P <0.5);(2) Decorin in the melatonin48,72hours,the concentration of1g/L the most significant effect (P<0.5).(3)OCIAD2genes belonged to cancer-related genesand the expression levels was relatively lower than actin, about0.This result suggested thatthe gene expression in normal cells was very low, the influence on its expression was notobvious.Bioinformatics methods and RT-PCR were used to analyse to the expressin of UMP/CMP kinase gene. The results showed that:(1)the UMP/CMP kinase gene is1081bp encodeda protein of228amino acid protein, showed a higher leucine, glycine, lysine content andlower content of leucine, tryptophan.The secondary structure prediction showed thatUMP/CMP kinase contained a higher proportion of random coil structure. Its protein domainswere analyzed and found to contain UMP/CMP kinase protein domain.(2) RT-PCR testresults show that the UMP/CMP kinase gene expression in the skin.Keratin associated protein was thought to affect the growth of the cashmere goat hair.Inorder to further study the physiological function of the protein, Liaoning new breed ofcashmere goat keratin associated protein KAP26.1was as the research objec for the proteinexpression, purification and preparation of polyclonal antibody. Results showed the proteinsassociated protein KAP26.1cloned to PET-32A prokaryotic expression vector in E. coli andachieved a high expression in the recombinant protein,which was purified as an antigen toimmunize rabbit, and preparing of rabbit anti-KAP26.1polyclonal antibody by ELISA method to measure the titer of the purified polyclonal antibody was obtained having a hightiter. Western blot results proved that antibodies can be prepared with keratin associatedprotein KAP26.1.The study made a theoretical foundation for further study of Liaoning new breed ofcashmere goat germplasm genetic characteristics. Research cashmere goat biological geneticcharacteristics, deepening our unique genetic resource-based theory and providing atheoretical basis from the molecular level to further protect the gene pool.