Effects Of N-3 PUFAs Diet On Obesity Mice And Construction Of Knock-In Vector For Fat-1 Transgenic Mice

Partâ… Objective To determine if n-3 PUFAs diet affected on mice obesity and obesity-related cell factors.Methods 36 healthy female mice were divided into three groups namely high fat diet group, n-3 PUFAs diet group and control group, to test the body weight and food intake weekly and the level of serum Adiponectin, Orexin-A and Resistin by ELISA.Results The adiposity and serum Orexin-A level in n-3 PUFAs diet model group were significantly lower than those in high fat diet model group (P<0.05), and the serum Resistin level in n-3 PUFAs diet model group was significantly higher than that in high fat diet model group (P<0.05).Conclusion There was an obvious reduce on the development of adiposity in mice when feed them n-3 PUFAs diet.Part IIObjective To construct the knock-in vector for polyunsaturated fatty acids dehydrogenase fat-1 gene cDNA site-direct homologous recombination into mice ES cell.Methods Two homologous arms flanking the target region obtained from mice genomic DNA (C57BL/6) by PCR was inserted into universal targeting vector pSSC-9, and expression cassette of this vector was improved through codon optimization, inclusion of a consensus Kozak sequence. Incorporated Cre-LoxP technology to enable neoR sequence can be eliminating after ES cell transfection.Results Our results show that correct structure of the knock-in vector was confirmed by restriction enzyme digestion and sequencing analysis, and demonstrate that a targeting vector for fat-1 gene site-direct knock-in has been successfully constructed.Conclusions The fat-1 transgenic mice knock-in vector could be use to transfect the embryonic stem cell of C57BL/6 mice.