The Genetic Analysis On HFJ And MIJ Rats With The Methods Of RAPD And SSR

Objective: The laboratory rat is one of the most important laboratory animals in experimental medicine and drug development, and inbred rat has its irreplaceable value also. There are about 1000 inbred strain rats now, but all of inbred rats in China are introduced from abroad. Technicians from Laboratory Animal Department of our university domesticated inbred rat from 2001.A pair of rats for both sex were chosen from a outbreed Wistar population which were breed in barrier environment, and two new inbred strains(HFJ and MIJ) with each characters formed initially via mating of brother-sister pairs for 20 generations.It has been observed that HFJ has higher fecundity than other inbred strains even outbreed strains. We could establish independent intellectual property rights of the new line of laboratory animals if we did further research and development.MIJ strain has testicular digenesis and genital system spontaneous tumor expressions and these expressions comply monogenic inheritance law. We can foster further development of animal model of spontaneous male sterility and animal model of reproductive system tumor as important tools for human diseases research and new drugs development. This strain can fill the blank in the world in spontaneous male sterility.There may be some genetic pollutions or genetic mutations companied by genetic characters changed because of human errors or animal's own variations when we breed the laboratory animals. The variations will impact the whole population, if we don't elimination the rats with variations. If these rats were used in some experiments, there will be a false result waiting for us. So it is necessary to make strictly , effectively monitor to keep the genetic stability. As national standards for detecting mammalian genetic stability, detections with biochemical markers and immunological markers are the most common methods. These methods don't detect genetic materials directly. The results may not be entirely consistent with the actual, because external performance is a interaction of genes and environmental. Molecular biological methods as supplement are necessary. So we introduce RAPDs and SSRs methods and compare them to find a better molecular biological method to be used for monitoring the rats'genetic stability.Method :1. Extraction of genomic DNAs Genomic DNAs for 2 rats for both sex of the 4 strains( HFJ,MIJ,Lewis,F344) were extracted with traditional phenol-chloroform extraction method and nucleic acid extraction instrument.2. Genetic reserch of 4 strains with RAPDs markers.60 RAPD primers bought from SBS Genetech were used to detect the genetic stabilities and diversities.3. Genetic research of 4 strains with microsatellite markers. 24 microsatellite primer pairs used to detect the genetic stabilities and diversities.Results:1. 12 of 60 RAPD primers were observed good amplifications. One primer C-19 can distinguish Lewis and F344 with HFJ and MIJ, another one M-10 can distinguish F344 with other strains .Polymorphism can't be observed between HFJ and MIJ or in each of the two strains.2. All of the 24 SSR primer pairs produced good amplified PCR products. Among the 24 primer pairs, 15(62.5%) turned out to be polymorphic. None significant deviation was observed between HFJ and MIJ. HFJ and MIJ strains have significant deviations from Lewis and F344, there is also significant differentiation between Lewis and F344.Two deviations were observed in HFJ, and two deviations were observed in MIJ. Another 11 HFJ rats and another 9 MIJ rats were investigated on thus loci, the result presented monomorphism.Conclusions:1. The characteristic RAPDs fingerprintings for HFJ and MIJ strains with 11 primers were set up. The two were same each other, but different with Lewis and F344.2. The characteristic SSR fingerprintings for HFJ and MIJ with 24 microsatellites loci were set up. There was little difference between the two strains, but there was significant difference between difference with Lewis and F344.3. HFJ and MIJ came from the same pairs of Wistar, so the gnomes of the two strains were very similar, but there is significant difference between difference with Lewis and F344.