基于猪细小病毒病毒样颗粒的结肠癌靶向纳米载体的构建

发布时间：2018-05-11 05:28

本文选题：TK肽 + 纳米载体　；参考：《微生物学通报》2017年10期

【摘要】：【目的】获得具有结肠靶向的纳米载体。【方法】采用SOE-PCR方法将具有结肠靶向的TK肽序列插入到猪细小病毒(PPV)结构蛋白VP2的环2和环4区域得到TK-vp2(?vp2)基因,在Bac-to-Bac?杆状病毒表达系统中构建、表达和自组装。【结果】通过SOE-PCR方法扩增获得?vp2基因,在Bac-to-Bac?杆状病毒表达系统中构建得到Bacmid-?vp2,经脂质体转染至Sf9昆虫细胞得到重组杆状病毒。直接免疫荧光试验、SDS-PAGE和Western blot检测结果表明?VP2蛋白在Bac-to-Bac?杆状病毒表达系统中获得融合表达,目的蛋白约70 k D;透射电子显微镜结果显示?VP2能自组装形成病毒样颗粒(TK-VLPs),直径范围在22 nm-30 nm。【结论】获得纳米载体TK-VLPs,为进一步研究其作为结肠靶向的纳米载体奠定物质基础。
[Abstract]:[objective] to obtain colon-targeting nanocarriers. [methods] the colon-targeted TK peptide sequence was inserted into the ring 2 and ring 4 region of porcine parvovirus (PPV) structural protein VP2 by SOE-PCR method, and the TK-vp2 / VP2) gene was obtained in Bac-to-Bac. The baculovirus expression system was constructed, expressed and self-assembled. [results] VP2 gene was amplified by SOE-PCR. Bacmid-VP2 was constructed from baculovirus expression system and transfected into Sf9 insect cells by liposome to obtain recombinant baculovirus. The results of SDS-PAGE and Western blot showed that VP2 protein was detected in Bac-to-Bac. Fusion expression was obtained in baculovirus expression system. The results of transmission electron microscopy (TEM) showed that VP2 could self-assemble into virus-like particles TK-VLPsN with a diameter range of 22 nm-30 nm. [conclusion] Nano-carrier TK-VLPswas obtained, which laid a solid foundation for further study of its role as colon-targeted nano-carrier.
【作者单位】：黑龙江八一农垦大学;哈尔滨医科大学;
【基金】：黑龙江八一农垦大学研究生创新科研资助项目(No.YJSCX2016-Y22) 黑龙江省自然基金-青年项目(No.QC2013C029) 哈尔滨医科大学伍连德基金(No.WLD-QN1111)~~
【分类号】：Q78;R73-36

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