Author: Zhang, XiZhen; Wang, XiaoDan; Zhao, DongHai; Meng, XiangYu; Zhao, XingHong; Yu, XiangHui; Kong, Wei
Title: Design and immunogenicity assessment of HIV-1 virus-like particles as a candidate vaccine Document date: 2011_12_16
ID: xxypq3wd_23
Snippet: To generate a stable cell line expressing our genes of interest, we optimized the ratio of vector D-GPEi and pcDNA3.1 and determined a 3:1 ratio was optimal. We changed the selective media (G418) after 48 h and selected for positive colonies that express the HIV-1 gag, pol and env structural proteins, as determined by Western blot assays (data not shown). To test for secreted protein, cell supernatant was collected by 30% sucrose density centrifu.....
Document: To generate a stable cell line expressing our genes of interest, we optimized the ratio of vector D-GPEi and pcDNA3.1 and determined a 3:1 ratio was optimal. We changed the selective media (G418) after 48 h and selected for positive colonies that express the HIV-1 gag, pol and env structural proteins, as determined by Western blot assays (data not shown). To test for secreted protein, cell supernatant was collected by 30% sucrose density centrifugation and pellets resuspended in PBS and assayed by SDS-PAGE and Western blot (Figure 1 ).
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