Vasculostatin慢病毒表达载体的构建

Construction of lentiviral vector expressing Vasculostatin

目的构建Vasculostatin慢病毒表达载体。方法通过基因合成方法获得Vasculostatin的完整序列,将此片段定向克隆到pL enti-CMV-EF1p-eG FP载体多克隆位点区,筛选重组质粒,采用磷酸钙方法将Lenti-CMV-Vasculostatin/EF1p-eG FP与载体pC MV-VSGS与pC MV-Gag-Pol共转染293T细胞,转染后收获上清离心纯化病毒并检测病毒滴度。结果成功构建Vasculostatin的慢病毒表达载体,检测病毒的滴度为2.5×107/ml。结论 Vasculostatin慢病毒表达载体为研究Vasculostatin对血管生成的抑制作用提供工具。

Objective To construct a lentiviral vector expressing Vasculostatin. Methods Vaseulostatin sequence was acquired by gene synthesis and was directional cloned into pLenti-CMV- EFI p-eGFP. Lenti-CMV-Vasculostatin/EFlp-eGFP was cotransfected with pCMV-VSGS and pCMV- Gag-Pol into HEK 293T cells, the supematant was collected to purified virus, and virus titers were determined. Results Lentiviral vector expressing Vasculostatin was constructed successfully and virus titers were 2.5× 107/ml. Conclusion Lentiviral vector expressing Vasculostatin can be used in studying the anti-angiogenic potent of Vasculostatin.