Studies On Dynamic Distribution And Expression Of GPV-VP3 Gene Vaccine In Mice

Studies on Dynamic Distribution and Expression of GPV-VP3 Gene Vaccine in Mice Liu Xiaodong (Biochemistry and Molecular Biology) Directed by Pro.Wang Mingshu , Pro.Cheng AnchunIn this thesis, research about the method of detecting VP3 gene of Gosling Plague Virus (GPV), the distribution and expression of GPV-VP3 gene vaccine and live attenuated GPV vaccine vaccinated into mice were as fellows:①the method of Fluorescent Quantitative PCR (FQ-PCR) for detecting VP3 gene of GPV was established;②GPV-VP3 gene vaccine (pcDNA-GPV-VP3) was vaccinated into six groups of BALB/c mice via intramuscularly injection with three dosage (200μg per mouse, 100μg per mouse and 50μg per mouse) and gene gun bombing with three dosage (6μg per mouse, 3μg per mouse and 1μg per mouse), live attenuated GPV vaccine was vaccinated into one group of BALB/c mice via intramuscularly injection with 100μl per mouse, then cardiac muscle, liver, spleen, lung, kidney, brain, intestine and tissues of immune site(muscle or cutis) were collected at intervals and established FQ-PCR and immune-histochemistry were used to detect the dynamic distribution and expression of pcDNA-GPV-VP3 and live attenuated GPV vaccine in these tissues. The results show that1. the method of FQ-PCR is specific, sensitive and good repeated, there was a good linear correlation between the copy numbers of the nucleic acid and Ct value detected by FQ-PCR (correlation coefficients= 0.999).2. pcDNA-GPV-VP3 could distribute into all the mice tissues and have the most. copy number 1h-3h post-inoculation, then the copy munber began to decrease, but pcDNA-GPV-VP3 could still been detected in all mice tissues 31 wk post-inoculation.3. in the three groups pcDNA-GPV-VP3 vaccinated via intramuscularly injection, there was positive correlation between the immune dosage and the copy numbers of gene vaccine, the difference among the three groups was significant before 15wk (P＜0.05) post- inoculationand not significant after 15wk (P＞0.05) post-inoculation; in the three groups pcDNA-GPV-VP3 vaccinated via gene gun bombing, there was weak positive correlation between the immune dosage and the copy numbers of gene vaccine, the difference among the three groups was notsignificant (P＜0.05).4. in the three groups pcDNA-GPV-VP3 vaccinated via intramuscularly injection, the expression products of pcDNA-GPV-VP3 were detected in nucleus of cardiac muscle cell, muscle cell of immune site, kidney cell and liver cell, there was some positive correlation between the immune dosage and the quantity of expression products ; in the three groups pcDNA-GPV-VP3 vaccinated via gene gunbombing the expression products of pcDNA-GPV-VP3 were detected in nucleus of cardiac muscle cell,cutis cell of immune site, kidney cell and liver cell, there was weak correlation between the immune dosage and the quantity of expression products.5. intramuscularly injection and gene gun bombing were both good methods for inoculation of pcDNA-GPV-VP3, the immune dosage of gene gun bombing was very small ,but in groups pcDNA-GPV-VP3 vaccinated via gene gun the expression products of pcDNA-GPV-VP3 could been detected first, the expression products were more and expressed longer, gene gun bombing was better than intramuscularly injection.6. GPV could distribute into all the mice tissues and have the most copy number 1 d-lwk post-inoculation, then the copy number decrease very fast but GPV could still been detected in all mice tissues 31 wk post-inoculation; GPV could been detected in nucleus of cardiac muscle cell, muscle cell of immune site, kidney cell and liver cell by immune-histochernistry.