| Alzheimer’s disease (AD) has became the fourth serious disease leading to the human death excepting for heart disease, cancer, and apoplexy. Previous study indicated that there were highly correlated both the expression of apolipoprotein E (ApoE) and the AD. Therefore, the objective of current study is to analyze the expression profiles of ApoE in different tissues, ages, and the different regions in brain in Guangxi Bama mini-pig. The eukaryotic expression vector of ApoE was constructed and the function of recombinant plasmid was identified. Meanwhile, the transgenic mice were conducted with the method of injection directly in ovaries and testes. The study will make preparation for the construction of animal model of AD in Guangxi Bama mini-pig.Specific primers were designed according to the sequence of ApoE gene released in GenBank using Oligo6software. The expression profiles of ApoE was analyzed by reverse transcription (RT) PCR in different tissues (heart, spleen, liver, kidney, pancreas, brain) and different ages (10,14,24,>24-month) in Guangxi Bama mini-pig. The results showed that the mRNA transcription levels of ApoE were significantly higher in brain and pancreas than that in heart, spleen, liver, and kidney. With the increasing of age, the expression levels of ApoE were elevated. The result of immunohistochemistry indicated that the expression levels of ApoE were obviously higher in forebrain cortex than that in hippocampus.ApoE cDNA was cloned and eukaryotic expression vector was constructed. The function of recombinant plasmid was transfect to PK-15and PC-12cells for identifying the function of plasmid. The results showed that there were the expressions of EGFP in PK-15and PC-12cells, which indicated that the recombinant plasmid could expressed in cells. The plasmids were injected into ovaries and testes of mice. A total of57F1generation mice were conducted by natural mating and two positive mice were identified by PCR. The transgenic rate was3.5%. |
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