中国组织工程研究与临床康复2008，Vol.12Issue(46)：9184-9188,5.
Der p 2基因真核表达载体构建及其在小鼠树突状细胞中的表达
Construction of eukaryotic expression vector of Der p2 gene and its expression in mouse dendritic cells
摘要
Abstract
BACKGROUND: Dendritic cells, the most potent antigen presenting cells known at present, have been extensively used in the immunotherapy as adjuvant. OBJECTIVE: The present study was to construct Der p 2 eukaryotic expression vector and validate its expression in the mouse bone marrow-derived dendritic cells. DESIGN, TIME AND SETTING: A single sample observation was performed at the Institute of Respiratory Disease, Xinqiao Hospital, Third Military Medical University of Chinese PLA between May and December 2005. MATERIALS: C57BL/6 mice were included. Plambd-Der p 2 was the product of Heska Company, USA.pCI-neo plasmid was provided by the Institute of Respiratory Disease, Xinqiao Hospital, Third Military Medical University of Chinese PLA.METHODS: Mouse bone marrow-derived dendritic cells were in vitro isolated and cultured.Complete Der p 2 cDNA was spliced from prokaryotic expression vector plambd-Der p 2, and then cloned into eukaryotic expression vector pCI-neo (pCI-neo-Der p 2).The positive recombinants pCl-neo-Der p 2 transfected into dendritic cells.Non-transfected and blank vector pCI-neo-transfected dendritic cells were used as controls. MAIN OUTCOME MEASURES: ①Identification of pCI-neo-Der p 2 recombinant plasmid.②Detection of Der p 2 mRNA and protein expression by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot techniques. RESULTS: Sequencing results showed Der p 2 cDNA in pCI-neo-Der p 2 was in coincidence with the sequence registrated in Gene Bank.RT-PCR and Western Blot results showed that expression of Der p 2 mRNA and protein could be detectable in the pCI-neo-Der p 2-transfected dendritic cells. CONCLUSION: The Der p 2 cDNA was successfully constructed into the eukaryotic expression vector, and Der p 2 gene and protein could be expressed efficiently in dendritic cells.关键词
Der p 2/载体构建/树突状细胞分类
医药卫生引用本文复制引用
毕玉田,王彦,吴奎,王长征,钱桂生..Der p 2基因真核表达载体构建及其在小鼠树突状细胞中的表达[J].中国组织工程研究与临床康复,2008,12(46):9184-9188,5.基金项目
国家自然科学基金资助项目(30470772) Supported by:the National Natural Science Foundation of China,No.30470772 （30470772）
