Human Neurobeachin-like 1(NBEAL1)gene was an important member of BEACH-WD40 domain family,which was confirmed to be overexpressed in I stage glioma.In this study,we extracted total RNAs from U251 cell line,acquired it...Human Neurobeachin-like 1(NBEAL1)gene was an important member of BEACH-WD40 domain family,which was confirmed to be overexpressed in I stage glioma.In this study,we extracted total RNAs from U251 cell line,acquired its cDNA sequence by RT-PCR,and cloned part of NBEAL1 cDNA fragments into the vector pGEX-KG.The recombinant expression vector achieved high expression in E.coli BL21 as a GST fusion protein.NBEAL1 recombinant protein was purified by affinity chromatography.Monoclonal antibody was prepared against the recombinant NBEAL1 protein.Its bioactivity was identified by Western Blotting analysis.Anti-NBEAL1 antibody was conjugated with CdTe quantum dots.Resultant anti-NBEAL1 antibody-conjugated nanoprobes were injected into mice via tail vessel.After 12h,it is clearly observed that prepared nanoprobes located in brain tissues of mice model with glioma by IVIS Imaging system.In conclusion,NBEAL1 protein was successfully expressed and its monoclonal antibody was successfully prepared.Anti-NBEAL1 antibody-conjugated quantum dots may be used to image glioma.These prepared nanoprobes have great potential in early detection of glioma.展开更多
基金supported by China National 973 Project(No.2010CB933901)National 863 Hi-tech Project(Nos.2007AA022004 and 2005AA001070)National Natural Scientific Fund(No.30672147).
文摘Human Neurobeachin-like 1(NBEAL1)gene was an important member of BEACH-WD40 domain family,which was confirmed to be overexpressed in I stage glioma.In this study,we extracted total RNAs from U251 cell line,acquired its cDNA sequence by RT-PCR,and cloned part of NBEAL1 cDNA fragments into the vector pGEX-KG.The recombinant expression vector achieved high expression in E.coli BL21 as a GST fusion protein.NBEAL1 recombinant protein was purified by affinity chromatography.Monoclonal antibody was prepared against the recombinant NBEAL1 protein.Its bioactivity was identified by Western Blotting analysis.Anti-NBEAL1 antibody was conjugated with CdTe quantum dots.Resultant anti-NBEAL1 antibody-conjugated nanoprobes were injected into mice via tail vessel.After 12h,it is clearly observed that prepared nanoprobes located in brain tissues of mice model with glioma by IVIS Imaging system.In conclusion,NBEAL1 protein was successfully expressed and its monoclonal antibody was successfully prepared.Anti-NBEAL1 antibody-conjugated quantum dots may be used to image glioma.These prepared nanoprobes have great potential in early detection of glioma.
