Fas和β-雌二醇受体融合基因在COS-7细胞中的表达
融合基因 序列分析 转染 免疫印迹
作者：孙 蕾1 王会信1 周廷冲1
单位：军事医学科学院基础医学研究所 北京 100850
关键词：融合基因 序列分析 转染 免疫印迹
免疫学杂志990103 摘 要 设计三条引物P1、P2、P3扩增全长hFas基因和只含有跨膜区和胞浆区的mFas基因。P2上设计一个BstXI位点，以与hFas信号肽之后的单酶切位点BstXI连接，获得带信号肽的跨膜区和胞浆区SpmFas基因。另外设计两条引物P4、P5扩增人β-雌二醇受体的激素结合结构域。扩增基因均经DNA序列分析得到证实。Ρ3、P4均设计了Kpn I位点，用于两基因融合，并将此融合基因插入真核表达载体pcDNA3。脂质体法转入COS-7细胞进行瞬时表达，Western Blot检测到约57KDa的表达产物，与融合基因的估算分子量相符。
中图号 Q78
EXPRESSION OF FAS/ESTROGEN RECEPTOR FUSION GENE IN COS-7 CELLS
Sun Lei， Wang Huixin， Zhou Tingchong
(Institute of Beijing Basic Medical Sciences， Beijing 100850)
Abstract Three primers P1、P2、P3 were used to amplify the whole sequence and the transmembrane and cytoplasma fraction of hFas by PCR. A BstX I site was designed in P2 to ligate with the BstX I site downstream hFas signal peptide to obtain SpmFas. Other two primers P4、P5 were used to PCR-amplify the hormone-binding domain of human β-estrogen receptor. All amplified fragments were verified by sequencing analysis. The two genes were fused at Kpn I site designed in both P3 and P4 and then the fusion gene was inserted into pcDNAs.Using lipofectamine-mediated gene transfer technique ......
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单位：军事医学科学院基础医学研究所 北京 100850
关键词：融合基因 序列分析 转染 免疫印迹
免疫学杂志990103 摘 要 设计三条引物P1、P2、P3扩增全长hFas基因和只含有跨膜区和胞浆区的mFas基因。P2上设计一个BstXI位点，以与hFas信号肽之后的单酶切位点BstXI连接，获得带信号肽的跨膜区和胞浆区SpmFas基因。另外设计两条引物P4、P5扩增人β-雌二醇受体的激素结合结构域。扩增基因均经DNA序列分析得到证实。Ρ3、P4均设计了Kpn I位点，用于两基因融合，并将此融合基因插入真核表达载体pcDNA3。脂质体法转入COS-7细胞进行瞬时表达，Western Blot检测到约57KDa的表达产物，与融合基因的估算分子量相符。
中图号 Q78
EXPRESSION OF FAS/ESTROGEN RECEPTOR FUSION GENE IN COS-7 CELLS
Sun Lei， Wang Huixin， Zhou Tingchong
(Institute of Beijing Basic Medical Sciences， Beijing 100850)
Abstract Three primers P1、P2、P3 were used to amplify the whole sequence and the transmembrane and cytoplasma fraction of hFas by PCR. A BstX I site was designed in P2 to ligate with the BstX I site downstream hFas signal peptide to obtain SpmFas. Other two primers P4、P5 were used to PCR-amplify the hormone-binding domain of human β-estrogen receptor. All amplified fragments were verified by sequencing analysis. The two genes were fused at Kpn I site designed in both P3 and P4 and then the fusion gene was inserted into pcDNAs.Using lipofectamine-mediated gene transfer technique ......
您现在查看是摘要页，全文长 9304 字符。