Use Of Somatic Cell In Bovine To Study The Expression Of The Related Fatty Acid Synthesis Genes

Fatty acids for the newborn animals a lot of energy and essential fatty acids, but high concentration of C14:0 (cardamom acid) and C16:0 (palmitic acid) can promote low-density lipoprotein cholesterol in plasma concentration,thereby inducing cardiovascular and cerebrovascular diseases on the potential health hazards.Therefore reasonable to improve the milk fat composition, to improve milk quality, milk production has been the subject focus of concern. For this purpose, the study was to make use of somatic cell in bovine to research on the expressions of the related fatty acid genes.To clarify the factors affecting milk fat synthesis and the intrinsic molecular mechanism to provide the theoretical basis. Especially in the nutritional regulation of milk fat and provide a theoretical basis for genetic improvement.?for the optimization of raw milk of ruminants.The results as follow:Preparation of milk somatic cell: Colostrum milk,normal milk and ending milk of Bovine was objection, Identificated the SCC by directoscope method,blood cell countion method and trypan blue exclusion staining method. And then extracted RNA from milk somatic cell according to the Puissant's method and determined the purity integrity and content of RNA by electrophoresis and UV spectrophotometer and housekeeping gene GAPDH by RT-PCR . The result show that: Blood cell count measured by SCC board about 10～18.3×104/ml;it proved the milk somatic cell taken from health cow.The trypan blue dye measured energy is about 60 ~ 67.5%, RNA OD260/OD280 is 1.65 ~ 2.0,the content is 0.6 ~ 1.0mg/ml. The result of RT- PCR show that specificity amplifyication strap of housekeeping gene GAPDH existed at 452bp and show that have been successfully prepared the milk somatic cell.Expressions of mammary gland fatty acid synthesis-related genes: Somatic cell in Colostrum milk,normal milk and ending milk was select as obuection and houseke- eping gene GAPDH was selected as intra-axxording,analysised LPL,CD36, VLDLR, ACSS2,ACSL1, FABP3,SCD,ACC,FASN,ADFP,XDH and BTN1A1 mRNA in milk by semi-quantitive RT-PCR. The results showed that LPL,CD36,VLDLR,ACSS2, ACSL1, FABP3,SCD,ADFP, XDH and BTN1A1 mRNA expressions of colostrum milk,normal milk and ending milk.The ACC, FASN mRNA expression only in the colostrum milk,normal milk and ending milk were not detected. The transcription lev- el of the relative quantitative results showed that normal milk and ending milk LPL, CD36,VLDLR,ACSS2, ACSL1,FABP3,SCD,ACC,FASN,ADFP, XDH and BTN1A1 mRNA compared with colostrum milk, the transcription level was significantly lower(P<0.05), normal milk compared ending milk the difference was not significant (P>0.05).?These results coincide with the cows lactation curve, sugges- ting that fatty acid synthesis dependent on lactation stage, colostrum milk synthesis was higher than that normal milk and ending milk period.