Studies On Expression Of Fatty Acid Desaturase In Transgenic Rabbits

In order to study on the genetic and expression of fatty acid desaturase transgenic rabbitsgenerated by intratesticular injection, Firstly we use PCR to detect the integration ofexogenous gene in transgenic rabbit of all the425, and then detect the relative content of thefatty acids by gas chromatography for all PCR-positive transgenic rabbits (182/267). At last,we analyze the fatty acids results by area normalization. All the n-6PUFAs and n-3PUFAsratio of F0， F1and F2generations were analyzed through SAS9.2program betweenexperimental and control group as well as the genetic expression of three generations,different ages and tissues. And the results are as follows:1、The comparison of fatty acid relative content between different tissues. There aremore saturated fatty acids in muscle than liver, and their n-6/n-3are not significant (P>0.05).The n-6/n-3of liver, muscles, ear tissue, kidneys and blood of the weaning are higher than the120days (P<0.05). Among these five of the weaning period, ear tissue of n-6/n-3is the lowest,and then muscle, blood, kidney and Liver. By analyzing the n-6/n-3of F0, F1and the weaningbetween muscle, liver and ear tissue, we found that individual transgenic effect can berepresented by any tissue.2、The n-6/n-3of transgenic rabbits between three generations all decrease comparedwith the control group. In the n-6PUFAs of F0transgenic rabbit’s muscle and liver, theC18:2n-6of experimental group is significantly lower than the control group (P <0.01). TheC18:3n-3of experimental group is significantly higher than the control group (P<0.05).C20:5n-3, C22:5n-3and C22:6n-3of liver are significantly higher the control group (P <0.01).C22:6n-3of muscle is significantly higher than the control group (P <0.05). The n-6/n-3of theexperimental group between muscle and liver are significantly lower than the control group(P<0.01), dropped from13.19and10.96to5.30and4.91. These results suggest thatexogenous of fatty acid desaturase gene plays its catalytic function in transgenic rabbiteffectively.The n-6/n-3of F1transgenic rabbit’ muscle and liver are significantly lower than thecontrol group (P <0.01), dropped from13.19and10.96to3.35and3.43. The n-6/n-3of F2ear tissue is significantly lower than the control group (P <0.01), dropped from2.87to1.06. These results suggest that exogenous of fatty acid desaturase gene can be passed on to thenext generation stably and effectively expressed.3、The comparison of n-6/n-3of muscle and liver between F0and F1shows that thereare no significantly difference (P>0.05). However, to liver, the n-6/n-3of F1are lower thanF0(P<0.01).4、The comparison of fatty acid relative content between different ages shows that then-6/n-3of the weaning between ear, liver and muscle are all significantly higher than the120days (P<0.01).According to the results above, n-3PUFAs can be synthesized from n-6PUFAs intransgenic rabbits catalyzed by exogenous gene Δ15which is stably inherited to the nextgeneration. It doesn’t work in weaning period, and maybe it’s caused by the age limitation.Besides, we found that individual transgenic effect can be represented by any tissues. We haveobtained67transgenic rabbits rich in n-3PUFAs, and it proved the efficiency and practicalityof technology system that generation of transgenic rabbit by testicular injection and detectionthrough PCR as well as gas chromatography.