ADIG Induce The Trans-differentiation Of Bovine Myoblasts

With the improving of the people’s life quality, more and more consumers are fond ofmarbling muscle, which has the meat quality of better tenderness and flavors, and anincrease in the amount and volume of the intramuscular fat cells can form ideal marblingmuscle. Therefore, it is a scientific problems to be solved, also a reserch hotspot in the beefcattle breeding to breed a new beef cattle variety which are rich in marbling muscle throughrevealing the mechanism of molecular regulation about the process of adipocytesproliferation and differentiation in bovine adipocytes, which can contribute to decrease thesubcutaneous fat and increase the intramuscular fat in current.The mRNA level of ADIG gene in fat cells were positively associated with the degree ofdifferentiation, and ADIG gene expression is closely related to the gene phenotype ofadipocyte, but the role of ADIG gene in the process of adipocyte differentiation is unclearnow. In order to investigate the functions of ADIG gene in the process of bovine myoblasttrandifferntiating into fat, we imported the Ad-ADIG recombinant adenovirus in bovinemyoblasts.The main results were as follows:1. Complete the preliminary purification of bovine myoblast by means of passaging, inwhich using cell medium containing horse serum (10%HS+20%FBS+20%DMEM).2. Complete the Ad-ADIG recombinant adenovirus concentrate propagation throughrecoverying freezed293cells and culturing in cell medium (10%FBS+10%DMEM).3. Ad-ADIG recombinant adenovirus infection efficiency in cattle myoblasts could reachto90%, and could induce bovine myoblasts transdifferentiate successfully. Real-timequantitative PCR detected that during the induced transdifferentiation process, ADIG genewas expressed at high levels in the0th,2nd,4th,6th,8th,10th,12th14thday,and lipid dropletsincreased significantly. The fat-related genes PPARγ, SREBPF1, and FAS relative expressionincreased in the process, and the muscle-related gene MyoD relative expression decreased inthe early.In summary, we complete the preliminary purification of bovine myoblast and the Ad-ADIG recombinant adenovirus concentrate propagation. In addition, We overexpressedADIG gene in bovine myoblast successfully and induced bovine myoblasts transdifferentiating with using Ad-ADIG recombinant adenovirus, which provides evidenceabout the role of ADIG gene in bovine adipose tissue during growth and development, andlays a theoretical basis for further study of the molecular mechanisms of bovine fatmetabolism and meat quality improvement.