Study On Microsatellite Instability On Chromosome1of Cef Infected With ALV-J

Although the studies on Microsatellite Instability (Microsatellite Instability, MSI) of cancer have been20years, there is rare reports about MSI in avian tumour at home and abroad except my laboratory using MSI studying on early diagnosis of Marek’s disease. Using microsatellite technology genomes changes of chicken embryos cells infected Avian leukemia Virus has not been reported. In this experiment, chicken embryos fibroblasts were infected by ALV-J virulent strain. Using53microsatellite markers on chicken chromosome1, it was to find whether chicken embryos fibroblasts microsatellite DNA changes, research the genome alteration the early stage of the virus infection and provide clues for the early diagnosis of disease and cancer of the molecular pathogenesis.There was23SPF chicken embryos tissue, DNA of each half of chicken embryos tissue was extracted. The other half tissue were used to culture chicken embryos fibroblasts (CEF), then CEF were infected with ALV-J for7days. The CEF were collected and DNA was extracted afer the ELISA test was positive. With the53microsatellite markers the motif of DNA from tissue and CEF were amplified. All the polymerase chain reaction products were tested by12%Polyacrylamide gel electrophoresis-silver stain.32microsatellite markers which showed different percentage of MSI accounted for60.37%of microsatellite markers. The highest MSI alteration percentage microsatellite ABR0169and ABR0522, the rate was52.17%and47.83%respectively. High frequency microsatellite instability MSI-H (MSI rate was more than20%) loci number was13, ABR0007, ABR0169, ABR0204, ABR0280, ABR0287, ABR0319, ABR0329, ABR0379, ABR0522, ADL0268, MCW0036, MCW0058, MCW0145, accounting for24.52%of chromosome1microsatellite markers.19loci, ABR0113, ABR0117, ABR0142etc, the rate of which was between4.35%and17.39%, showed a low frequency microsatellite instability MSI-L. MSI of21loci did not occer.Microsatellite instability (MSI) was found in23samples out of23. The sensitivity of4samples,19(28.30%),22(26.42%),14(24.53),15(20.75%), were up to20.00%. The rate of half of all samples was between10%-20%, such as1,2,3, etc. MSI rate of only seven samples was under10%.The microsatellite instability was found in chromosome1of chicken embryo fibroblast infected with ALV-J. There was widely MSI on the early stage of tumorigenesis with a lot of research reports. This thesis indicated the same result. The microsatellite instability in the assy maye be suggested as a valuable method for the early stages of infection with ALV, and the further study of the humuan pathogenesis and immune control of virus-induced lymphoma.