【摘要】 通过多孔CaCO3微粒吸附伪狂犬病毒(Pseudorabies,PRV)基因组DNA,并在其表面依次交替聚合PLL、Alg至7层;以EDTA溶解去除CaCO3内核,制成聚PLL/Alg包被DNA的载体并感染家兔,观察PRV基因组DNA在体内的复制情况。结果显示,Na2CO3与CaCl2反应可获得直径4～6μm的多孔CaCO3微粒,对DNA的吸附效率约为1mg CaCO3微粒可吸附1μg DNA。经PLL/Alg包被后获得直径1～2μm、囊膜厚约200nm的PRVDNA聚PLL/Alg微囊。肌肉注射含6.5μg PRV DNA的PLL/Alg微囊,可引起试验兔死亡,经PCR检测确定为PRV感染引起的。结果表明,聚PLL/Alg微囊能够介导DNA高效转染,在作为DNA疫苗载体方面具有良好应用前景。更多还原

【Abstract】 To investigate the efficiency of poly-PLL(Poly-L-lysine)/Alg(Alginate) vector-mediated virus genomic DNA transfection and the virus genomic DNA’s biological activity in vivo.After PRV genomic DNA adhered to the porous CaCO3 particles,PLL and Alg were alternately polymerized on the surface of the porous CaCO3 particles to seven layers;then dissolve it with EDTA to remove CaCO3 core,get the vector in which the DNA coated by poly-PLL/Alg,infect the rabbits,and observe the replication of viral DNA.The result indicated that porous CaCO3 particles,4-6 μm in diam,were obtained from the reaction between Na2CO3 and CaCl2.Their efficiency of absorbing DNA was 1 mg/g CaCO3 particles.After coated by PLL/Alg,PRV DNA poly-PLL/Alg microcapsules were obtained,1-2 μm in diam,about 200 nm in capsules’ thickness.6.5 μg of PRV DNA microcapsules coated by PLL/Alg could cause rabbit death by intramuscular injection.It had been identified by PCR that the death was caused by PRV infection.In conclusion,poly-PLL/Alg microcapsules can mediate efficient transfection of DNA and have good prospects for the application in the area of DNA vaccine vectors.更多还原