Genetic Characteristics And Functional Verification Of Bovine FABP3 Gene In Transgenic Mice

The FABP3 gene is one of the important genes that influence the intramuscular fat deposition in beef cattle. It is positively related with marbling, back fat and shear force. Therefore it is transferred into mouse to study genetic characteristics and expression of bovine FABP3 gene in transgenic mice, which plays a key role on culturing the breeds that produced higher yield sand improved quality by transgenic methods. The genetic characteristics and expression of bovine FABP3 in transgenic mice were investigated in this study. The article mainly transfers the cattle FABP3 into the mouse, and detects the gene positive and the genetic in the generation. Meamwhile, real-time PCR and tissue in sutu expression verification the exogenous gene in the mice canbe expressed. The nutrition was immpact the mice weight and fat metabolism related blood biochemical significant. It is proved that the mouse carried the cattle FABP3 gene enable to gentic and express. The foundation will establish in the transgenic bovime, and the theory basis for molecular genetic breeding. The results of the research have shown follews:(1)By means of the living fluorescent identification and RT-PCR detection, the results showed that the four positive transgenic mice(2 male and 2 female) were identified as the positive transgenic mice, which could be used in the further genetic detection and functional analysis.(2)Four scenarios of mating were carried out to obtain the G1 generation. Amplified fragment by sequencing and random blasted, it can be seen that the amplified fragment of positive individual all contained the EGFP vector and the cattle FABP3 gene; Among that, the averagely positive rates of G1 is 67.8%. Apart from that, there exist difference between different groups and the highest is 100% in group 1.(3)G1 generation has 100% positive group, the experimental animalsin the group have random mating and extend to the G4 generation. PCR amplification and Blast compere verify the positive. The results shows G2 generation have 100% positive rate. G3 generation have 33.3% positive rate. G4 generation have 18.75% positive rate. It canbe proved that transgenic mice enable genetic to the next generation. But wth the increase of generation time, the purpose gene in progeny individual’s ratio decreasing.(4)Real time PCR detect the G0,G1 generation transgenic mouse FABP3 gene expression in the myocardial and skeletal muscle on mRNA level. The results indacate the transgenic mice total FABP3 have a significant difference expression in the myocardial and skeletal muscle compare with the wild type.(5)Western blot detect G0、G1、G2、G3 generation GFP protein expression in the transgenic mice myocardial and skeletal muscle. And the total FABP3 protein of mouse canbe detected. The results shown that transgenicmouseFABP3 is higher than wide type mouse. GFP protein has expressed in the transgenic mice myocardial and skeletal muscle.(6)The feeding tests were conducted for G1 generation. That is the validation for the function of FABP3 gene under the high oil diet conditions. The result shows that the weight of transgenic mice is not significantly different with that of wild mice in high oil diet group and normal diet group.Most importantly, the transgenic mice with high oil diet is significantly different than that of wild mice with normal diet(P<0.05). Through the detection of chemical indicator in blood sample, the content of Cholesterol, Triglyceride, High density lipoprotein, Low density lipoprotein, Creatinine are significantly higher that that of control mice.(7)Transgenic mice G0, G1 generation of individual expression in situ testing, in G0 generation individuals in higher quantity, G1 generation individuals can see green fluorescence, but there has slightly reduced. The results also showed that GFP protein can be expressed in mice, and canbe pass on to the next generation. The result is consistent with GFP protein detection in western blot.(8)By the means of gene capture, the integration site of exogenous gene in mouse genome was analyzed. The results showed that the two insertion site of exogenous gene located on Chr. 9 and Chr. 12, and it contained the repetitive sequence.