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学科主题: 临床医学
题名:
Gene cloning of secreted VEGF receptor flt-1 (I similar to IV extracellular domain) and the corresponding recombinant adenovirus construction and expression
作者: Zhang, H; Shou, CC
关键词: adenovirus ; flt-1 ; homogenous recombination ; expression
刊名: PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
发表日期: 2002-08-01
卷: 29, 期:4, 页:625-630
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemistry & Molecular Biology ; Biophysics
研究领域[WOS]: Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]: TRANSDUCTION
英文摘要:

The increasing data show that the soluble form of Flt-1 have the ability to block the biological activity of VEGF due to its high affinity to VEGF and formation of homologous. and heterogenous dimmer with transmembrane Flt-1 and KDR. The binding activity of Flt-1 focuses on its II and III extracellular domain. cDNA fragments encoding for signal peptide and flt-1 I similar to IV extracellular domain were amplified through RT-PCR with a pair of specific primers from human umbilical vein endothelial cells. Then, the sflt-1 gene was subcloned into pcDNA3-EF-1 vector, and the expression unit of EF-1-fit-1 was subcloned into shuttle vector of pAdTrack-CMV, linearized pAdTrack-EF-1-flt-1 was co-transformed into BJ5183 cells with adenoviral genomic plasmid of pAdEasy-1. The identified recombinant DNA was transfected into 293 cells to package adenovirus. From the supernatant and cell lysis, the presence of recombinant adenovirus was proved by PCR, and RT-PCR detection demonstrated the transcription of flt-1 in MGC803 cells infected with Ad-Track-EF-1-flt-1, and immunoprecipitation reveals that soluble Flt-1 can be secreted into the culture supernatant of infected MGC803 cells. These results indicated that tumor cells infected with the prepared recombinant adenovirus vector Ad-Track-EF-1-flt-1 can secrete soluble Flt-1, which will be further used for in vivo antiangiogenesis experiments.

语种: 中文
WOS记录号: WOS:000177560000026
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/68009
Appears in Collections:北京大学临床肿瘤学院_期刊论文

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作者单位: Peking Univ, Sch Oncol, Beijing Inst Canc Res, Beijing 100034, Peoples R China

Recommended Citation:
Zhang, H,Shou, CC. Gene cloning of secreted VEGF receptor flt-1 (I similar to IV extracellular domain) and the corresponding recombinant adenovirus construction and expression[J]. PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS,2002,29(4):625-630.
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