鸡IFN-γ DNA壳聚糖纳米粒的制备及外源基因表达

Preparation of chicken gamma-IFN DNA-Chitosan nanoparticles for gene expression in vitro

为检测纳米载体对表达重组质粒的保护效果,本实验构建了鸡γ干扰素(IFN-γ)真核表达重组质粒pCAGGS-ChIFN-γ;并将其与200μg/mL壳聚糖醋酸溶液制备成DNA/壳聚糖纳米颗粒(CNPs)。透射电镜观察CNPs呈不规则球形,光子相关色谱(PCS)仪测定显示CNPs的平均粒径为172.6nm±5.1nm;Zeta电位为20.8mV±2.9mV;CNPs重组质粒包埋效率和载药量分别为91.21%±2.54%和31.93%±0.16%。鸡胚成纤维细胞(CEF)体外转染实验表明:携带的外源基因能在CEF中释放、表达,其表达效率约为阳离子脂质体转染试剂LipofectamineTM 2000转染后的16.3%,同时CNPs对细胞基本无毒性。CNPs稳定实验也证实其稳定性良好。

In present study,the protection of nanoparticles on recombinant plasmid was investigated by evaluating the DNA-chitosan nanoparticles（CNPs） carrying chicken interferon-γ（ChIFN-γ） recombinant plasmid.The nanoparticles were spherical shape with a mean diameter of 1726 nm under the electronic microscope and had the Zeta potential of 20.8 mV ±2.9 mV measured by the Photon Correlation Spectroscopy（PCS）.The encapsulation efficiency of DNA was 91.21 %±2.54 % and the DNA content in the nanoparticles was 31.93 %±0.16 %.The capsulated DNA could be protected from the degradation by DNaseⅠ.The transfection efficiency of CNPs was about 16.3% of that the LipofectamineTM 2000 reagent.Our results also showed that CNPs could express successfully in the CEF cells,and were nontoxic to cultured cells.