• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>BMC Biotechnology >Purification of functional baculovirus particles from silkworm larval hemolymph and their use as nanoparticles for the detection of human prorenin receptor (PRR) binding
【24h】

Purification of functional baculovirus particles from silkworm larval hemolymph and their use as nanoparticles for the detection of human prorenin receptor (PRR) binding

机译：从家蚕幼虫血淋巴中纯化功能性杆状病毒颗粒并将其用作检测人肾素原受体（PRR）结合的纳米颗粒

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Background Baculovirus , which has a width of 40 nm and a length of 250-300 nm, can display functional peptides, receptors and antigens on its surface by their fusion with a baculovirus envelop protein, GP64. In addition, some transmembrane proteins can be displayed without GP64 fusion, using the native transmembrane domains of the baculovirus. We used this functionality to display human prorenin receptor fused with GFPuv (GFPuv-hPRR) on the surface of silkworm Bombyx mori nucleopolyhedrovirus (BmNPV) and then tested whether these baculovirus particles could be used to detect protein-protein interactions. Results BmNPV displaying GFPuv-hPRR (BmNPV-GFPuv-hPRR) was purified from hemolymph by using Sephacryl S-1000 column chromatography in the presence of 0.01% Triton X-100. Its recovery was 86% and the final baculovirus particles number was 4.98 × 108 pfu. Based on the results of enzyme-linked immunosorbent assay ( ELISA ), 3.1% of the total proteins in BmNPV-GFPuv-hPRR were GFPuv-hPRR. This value was similar to that calculated from the result of western blot by a densitometry (2.7%). To determine whether BmNPV-GFPuv-hPRR particles were bound to human prorenin, ELISA results were compared with those from ELISAs using protease negative BmNPV displaying β1,3- N -acetylglucosaminyltransferase 2 fused with the gene encoding GFPuv (GGT2) (BmNPV- CP --GGT2) particles, which do not display hPRR on their surfaces. Conclusion The display of on the surface of the BmNPV particles will be useful for the detection of protein-protein interactions and the screening of inhibitors and drugs in their roles as nanobioparticles.
机译：背景杆状病毒的宽度为40 nm，长度为250-300 nm，通过与杆状病毒包膜蛋白GP64融合，可以在其表面展示功能性肽，受体和抗原。另外，使用杆状病毒的天然跨膜结构域，可以显示一些跨膜蛋白而无需GP64融合。我们使用此功能来展示与GFP uv （GFP uv -hPRR ）在蚕Bombyx mori核多角体病毒（BmNPV）的表面上，然后测试这些杆状病毒颗粒是否可用于检测蛋白质-蛋白质相互作用。结果显示BmNPV显示GFP uv -hPRR（BmNPV-GFP uv -hPRR）是从在0.01％Triton X-100存在下，使用Sephacryl S-1000柱色谱分析血淋巴。回收率为86％，杆状病毒最终颗粒数为4.98×10 8 pfu。根据酶联免疫吸附试验（ELISA）的结果，BmNPV-GFP uv -hPRR中总蛋白的3.1％为GFP uv -hPRR。该值类似于通过光密度法从蛋白质印迹的结果计算出的值（2.7％）。为了确定BmNPV-GFP uv -hPRR颗粒是否与人肾素结合，将ELISA结果与使用β1,3-N的蛋白酶阴性BmNPV的ELISA结果进行了比较。 -乙酰氨基葡萄糖氨基转移酶2与编码GFP uv （GGT2）的基因融合（BmNPV- CP - -GGT2）粒子，这些粒子在其表面上不显示hPRR。结论BmNPV颗粒表面的展示可用于检测蛋白质与蛋白质的相互作用，以及筛选抑制剂和药物作为纳米生物颗粒的作用。

著录项

相似文献

  • 外文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号