Construction and identification of mouse R-spondin 1 eukaryotic expression vector
Construction and identification of mouse R-spondin 1 eukaryotic expression vector
复制标题
小鼠R-spondin 1真核表达载体的构建及鉴定
DOI：
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发表时间：
2013
影响因子：
--
通讯作者：
Hua J
中科院分区：
文献类型：
--
作者：
Peng S;Han W;Yao X;Hua J
R-spondin 1（Rspo 1）被认为是激活Wnt信号通路并与之协同作用。它参与胰腺和肌肉发育、骨形成、肢体和性腺形态发生，甚至与许多人类疾病有关。本研究利用RT-PCR技术从小鼠胰腺组织中扩增了Rspo 1基因的全长。将pMD 19-T载体与Rspo 1全长基因连接，构建重组载体pMD 19-T-Rspo 1，将测序正确的Rspo 1全长片段经EcoR I和BamH I双酶切后连接到pIRES 2-AcGFP 1载体上。重组质粒pRspo 1-IRES 2-AcGFP 1经EcoR I和BamH I双酶切鉴定。转染HEK 293细胞48 h后，荧光显微镜下观察绿色荧光蛋白（GFP）的表达，RT-PCR检测Rspo 1 mRNA的表达水平。结果表明，Rspo 1真核表达载体构建成功，并得到鉴定。这为进一步研究Rspo 1对Wnt信号通路的调控功能及对干细胞增殖分化的作用机制奠定了基础。
R-spondin1 (Rspo1) has been proposed to activate and synergize with the Wnt signaling pathway. It is involved in pancreas and muscle development, bone formation, limb and gonad morphogenesis, even associated with many human diseases. In this study, we amplified the full-length Rspo1 gene from mouse pancreas using RT-PCR technology. The recombinant pMD19-T-Rspo1 vector was constructed by connecting pMD19-T vector and full-length Rspo1 gene, and then the Rspo1 full-length fragment sequenced correctly was connected to pIRES2-AcGFP1 vector after double digestion with EcoR I and BamH I. The recombinant vector pRspo1-IRES2-AcGFP1 was identified by double digestion of EcoR I and BamH I. We examined the expression of green fluorescent protein (GFP) under fluorescent microscope and the mRNA expression level of Rspo1 using RT-PCR after transfection it into HEK293 cells for 48 h. These results indicated that Rspo1 eukaryotic expression vector had been constructed successfully and identified. This would lay the foundation for further research on regulatory function of Rspo1 on Wnt signaling pathways and mechanism on proliferation and differentiation of stem cells.
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影响因子：
3.3
作者：
Kim, Kyung-Ah;Wagle, Marie;Abo, Arie
通讯作者：
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影响因子：
3.5
作者：
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通讯作者：
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影响因子：
4.3
作者：
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通讯作者：
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影响因子：
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作者：
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影响因子：
4.7
作者：
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通讯作者：
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