[Construction of recombinant vector pEGFP-ANG and expression of angiogenin gene in HUVECs].
OBJECTIVE
To express the recombinant angiogenin protein in mammalian cells.


METHODS
Human ANG full-length cDNA was obtained by chemical synthesis. The target gene ANG was inserted into eukaryotic expression vector pEGFP-C2. The recombinant plasmid was transfected into the human umbilical vein endothelial cells (HUVECs) via Lipofectin transfection. The expression of ANG gene in transfected HUVECs was detected by fluorescence microscopy and immunohistochemical staining.


RESULTS
DNA sequencing showed the sequence of the synthetic ANG was correct and amino acid sequence of ANG was right. The green fluorescence could be seen in transfected HUVECs under fluorescence microscope. Immunohistochemical staining detection showed that ANG expressed in transfected cells.


CONCLUSIONS
Human ANG full-length cDNA has been obtained. The ANG protein was expressed in mammalian cells successfully.
Zhang
Hong-bin
HB
Department of Medical Research, General Hospital of Guangzhou Military Command, Guangzhou 510010, China. zhangwater@163.net
Wang
Jie
J
Wu
Jie
J
Jiang
Yue-hua
YH
Yang
Tai-cheng
TC
Xian
Jiang
J
Yang
Chuan-hong
CH
Zheng
Wen-lin
WL
D000595:Amino Acid Sequence; D001483:Base Sequence; D018076:DNA, Complementary / Q000737:chemistry; D042783:Endothelial Cells / Q000378:metabolism*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D012259:Ribonuclease, Pancreatic / Q000235:genetics*; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology
D018076:DNA, Complementary; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C046366:angiogenin; D012259:Ribonuclease, Pancreatic