聚赖氨酸-海藻酸微囊介导的病毒基因组DNA转染  

Poly-lysine-alginate microcapsules mediated virus genomic DNA transfection

作　　者：耿莉[1] 杨盛[1] 张丹[2] 高婷婷[1] 王国卿[1] 张乃生[3] 

机构地区：[1]东北大学生物技术研究所,辽宁沈阳110004 [2]南加州大学维特比工程学院,加利福尼亚洛杉矶90089 [3]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医学报》2013年第7期1051-1054,1107,共5页Chinese Journal of Veterinary Science

基　　金：中央高校基本科研业务费资助项目(N110405008)

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。To investigate the efficiency of poly-PLL（Poly-L-lysine）/Alg（Alginate） vector - media- ted virus genomic DNA trans{ection 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 pol- ymerized on the surface of the porous CaCO3 particles to seven layers lthen dissolve it with EDTA to remove CaCO3 core,get the vector in which the DNA coated by poly-PLL/Alg,infect the rab- bits, and observe the replication o{ viral DNA. The result indicated that porous CaCO3 particles, 4- 6 ;m in diarn,were obtained from the reaction between Na2 CO3 and CaC12. Their efficiency of ab- sorbing DNA was 1 mg/g CaCO3 particles. After coated by PLL/Alg, PRV DNA poly-PLL/Alg microcapsules were obtained, 1-2 t;m in diam,about 200 nm in capsules＇ thickness. 6.5 ;g of PRV DNA microcapsules coated by PLL/AIg 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 applica- tion in the area of DNA vaccine vectors.

关 键 词：多聚赖氨酸-海藻酸 微囊 基因组DNA 转染 

分 类 号：Q78[生物学—分子生物学] S852.23[农业科学—基础兽医学]