[1]安晓静,钱桂生,王长征,等.小鼠IDO真核表达载体的构建及其在未成熟树突状细胞中的表达[J].第三军医大学学报,2008,30(11):1033-1036.
 AN Xiao-jing,QIAN Gui-sheng,WANG Chang-zheng,et al.Construction of eukaryotic expression vector containing mouse IDO gene and its expression in immature DCs[J].J Third Mil Med Univ,2008,30(11):1033-1036.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第11期
页码:
1033-1036
栏目:
论著
出版日期:
2008-06-15

文章信息/Info

Title:
Construction of eukaryotic expression vector containing mouse IDO gene and its expression in immature DCs
作者:
安晓静钱桂生王长征夏俊波党涛廖伟
第三军医大学新桥医院:全军呼吸内科研究所,全军呼吸病研究重点实验室;全军心血管病研究所
Author(s):
AN Xiao-jing QIAN Gui-sheng WANG Chang-zheng XIA Jun-bo DANG Tao LIAO Wei
Institute of Respiratory Diseases,Department of Cardiovascular Diseases, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China
关键词:
吲哚胺23-双加氧酶绿色荧光蛋白真核表达载体未成熟树突状细胞
Keywords:
indoleamine 23-dioxygenase green fluorescent protein eukaryotic expression vector immature DC
分类号:
R338.12; R394-33; R394.2
文献标志码:
A
摘要:
目的 构建小鼠pEGFP-N1-IDO真核表达载体并观察其在未成熟树突状细胞中的表达。  方法  利用RT-PCR方法扩增小鼠IDO基因全长,利用DNA重组技术将其定向插入到真核表达载体pEGFP-N1。经酶切和测序鉴定后, 用DOTAP脂质体转染法转染未成熟树突状细胞。通过G418筛选, 用倒置荧光显微镜观察转染的未成熟树突状细胞绿色荧光蛋白的表达,用RT-PCR、Western blot检测IDO的表达。  结果  小鼠全长IDO基因序列正确插入pEGFP-N1载体,与GenBank中报道的序列一致,成功构建了pEGFP-N1-IDO真核表达载体,并转染未成熟树突状细胞,成功地表达目的基因。  结论 真核表达载体成功构建和转染未成熟树突状细胞,并证明能有效表达于未成熟树突状细胞中。
Abstract:
Objective    To construct a eukaryotic expression vector containing mouse IDO gene fused with enhanced green fluorescent protein (pEGFP-N1) and its expression in immature DCs.     Methods    The full-length IDO gene was obtained from mouse by RT-PCR and was inserted into eukaryotic expression vector pEGFP-N1. After identified by restriction digestion and sequencing, the recombinant pEGFP-N1-IDO was transfected into immature DCs by DOTAP liposome. After screened by G418, the green fluorescence protein expression in immature DCs was observed under inverted fluorescence microscope and the mRNA and protein expressions of IDO were respectively detected by RT-PCR and Western blot.      Results    The full-length sequence of IDO was successfully inserted into the eukaryotic expression vector pEGFP-N1 and confirmed by restriction digestion and sequencing. The recombinant was successfully transfected into immature DCs and the IDO gene was expressed.     Conclusion    The eukaryotic expression vector pEGFP-N1-IDO was constructed successfully and IDO gene could be expressed efficiently in immature transfected DCs.

参考文献/References:

安晓静,钱桂生,王长征,等.  小鼠IDO真核表达载体的构建及其在未成熟树突状细胞中的表达[J].第三军医大学学报,2008,30(11):1033-1036

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更新日期/Last Update: 2008-06-06