Research On Salusin-a Gene Of Eukaryon Expression Vector Constuction And Expression In HEK293 Cells

OBJECTIVE: To study the eukaryotic green fluorescent protein expression vector Salusin-a(pEGFP-Salusin-a) plasmid construction and its expression in HEK293 cells. To clarify the relationship between serum levels of salusin-a and the coronary atherosclerosis.METHODS: The serum salusin-a concentration of 92 patients underwent coronary angiography after admisson were assayed by ELISA. Characteristics and the relationship between salusin-a and the elements were studied in each group.We amplified the Salusin-a gene by RT-PCR, and cloned into multiple clone sites of pEGFP-N3 vector. The pEGFP-Salusin-a recombinant plasmids were digested by restriction enzyme, bacterial-colony PCR and sequencing were used for identifying. The efficiency of pEGFP-Salusin-a transfected into HEK293 cells by liposome was checked. The HEK293 cell were then randomly divided into three groups: control group, empty plasmid transfection group, and recombinant plasmid tansfection group. The Salusin-a gene expression in HEK293 cells was analysised by fluorescence intensity and RT-PCR.RESULTS: Salusin-a levels in the coronary artery disease groups were lower than controls.Serum salusin-a levels were reduced accordance with the Gensini score.The pEGFP-Salusin-a plasmid was successfully constructed. The efficiency of pEGFP-Salusin-a transfected into HEK293 cells was about 86%. And we found that both fluorescence intensity and Salusin-a mRNA of empty plasmid transfection group and recombinant plasmid tansfection group were significant higher than the control group.CONCLUSIONS:Serun salusin-a levels were significantly lower in coronary artery disease patients. The pEGFP-Salusin-a can be transfected into HEK293 cells and expressed, which provide a experimental base to research the Salusin-a gene function.