目的研究采用表面等离子共振(SPR)技术,检测不同相对分子质量壳聚糖对质粒DNA(pDNA)的保护作用,以防止核酸酶(DNaseⅠ)的降解。方法用具有羧基化葡聚糖表面的CM5芯片固定在Biacore3000生物大分子相互作用系统中制成壳聚糖芯片,采用SPR检测在核酸酶存在下,不同相对分子质量壳聚糖对pDNA的保护作用。结果在人体温度(37℃)下,浓度5U/μ1的DNaseⅠ在lmin内即可降解没有保护的质粒DNA,而相对分子质量为200000以上的壳聚糖对质粒具有良好的保护作用。结论表面等离子共振技术可以作为体外研究金属表面高分子对于质粒DNA保护作用的一种新方法。
Objective To study the protective effect of various molecular weights (MW) chitosan on plasmid DNA (pDNA) by surface plasmon resonance (SPR) technique,in order to prevent from nuclease degrading. Methods The SPR was performed to evaluate the effects of various MW ehitosan samples on the protection of pDNA from nuclease degradation. Results The results showed that the pDNA was completely degraded within 1 minute in 5 U/μl ofdeoxyribonuclease (DNase Ⅰ ) solution at 37 ℃. While the chitosan with MW more than 200 000 showed favourable protective effect on DNA. Conclusion The SPR could become a novel approach to study the protective effect of maeromoleeular substances with metallic surface on pDNA.