A novel alternatively spliced form of murine vascular endothelial growth factor, VEGF 115

被引:40
作者
Sugihara, T
Wadhwa, R
Kaul, SC
Mitsui, Y
机构
[1] Agcy Ind Sci & Technol, Natl Inst Biosci & Human Technol, Tsukuba, Ibaraki 305, Japan
[2] Chugai Res Inst Mol Med, Niihari, Ibaraki 30041, Japan
关键词
D O I
10.1074/jbc.273.5.3033
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Murine immortal fibroblasts express a form of vascular endothelial growth factor (VEGF) that was cloned, characterized and named VEGF 115, It differs from VEGF 120 by 37 amino acids at the carboxyl terminus, VEGF 115-specific sequence reacted to a single transcript in mouse tissues, Reverse transcription-polymerase chain reaction was performed in mouse tissues and in fibroblasts of normal and immortal divisional phenotypes. The data from mouse tissues suggested that VEGF 115 is not a tissue-specific isoform of VEGF 120, whereas a functional relevance with immortalization is indicated from the latter, The novel cDNA was expressed in Escherichia coli, and the His-tagged VEGF 115 (17.2 kDa) thus obtained was recognized by anti-VEGF antibody, A mammalian expression plasmid, pCMV(neo+), encoding for VEGF 115 was transfected to NIH 3T3 cells, and the conditioned medium of stable transfectants was found to have fibroblast growth factor-replacing activity for human umbilical vein endothelial cells. Two independent genomic PI clonings with primers specific for VEGF 164 and VEGF 115, respectively, resulted in isolation of identical P1 clones, We analyzed these three P1 clones on Southern blots with common and specific probes for VEGF 164 and VEGF 115. The results support the hypothesis that VEGF 115 is a new alternatively spliced form of mouse VEGF.
引用
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页码:3033 / 3038
页数:6
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