지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2012.6
- 수록면
- 160 - 164 (5page)
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The baculovirus/insect cell expression system is of great value for the large-scale production of normal and mutant mammalian passive glucose-transport proteins heterologously for structural and functional studies. In most mammalian cells that express HepG2, this transporter isoform is predominantly located at the cell surface. However, it had been reported that heterologous expression of other membrane proteins using the baculovirus system induced highly vacuolated cytoplasmic membranes. Therefore, how a cell responds to the synthesis of large amounts of a glycoprotein could be an interesting area for investigation. In order to examine the subcellular location of the human HepG2 transport proteins when expressed in insect cells, immunofluorescence studies were carried out. Insect cells were infected with the recombinant baculovirus AcNPVHIS-GT or with wild-type virus at a MOI of 5, or were not exposed to viral infection. A high level of fluorescence displayed in cells infected with the recombinant virus indicated that transporters are expressed abundantly and present on the surface of infected Sf21 cells. The evidence for the specificity of the immunostaining was strengthened by the negative results shown in the negative controls. Distribution of the transporter protein expressed in insect cells was further revealed by making a series of optical sections through an AcNPVHIS-GT-infected cell using a confocal microscope, which permits optical sectioning of cell sample. These sections displayed intense cytoplasmic immunofluorecence surrounding the region occupied by the enlarged nucleus, indicating that the expressed protein was present not only at the cell surface but also throughout the cytoplasmic membranous structures.
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감사의 글
REFERENCES
참고문헌 (24)
참고문헌 신청
Asano T / 1989 / Rabbit brain glucose transporter responds to insulin when expressed in insulin-sensitive Chinese hamster ovary cells / J Biol Chem 264:3416~3420
Bell GI / 1993 / Structure and function of mammalian facilitative sugar transporters / J Biol Chem 268:19161~19164
Cope DL / 1994 / Domain assembly of the GLUT1 glucose transporter / Biochem J 300:291~294
Germann UA / 2000 / Expression of the human multidrug transporter in insect cells by a recombinant baculovirus / Biochemistry 29:2295~2303
Gorga FR / 1981 / Equilibria and kinetics of ligand binding to the human erythrocyte glucose transporter. Evidence for an alternating conformation model for transport / Biochemistry 20:5108~5113
Bell GI / 1993 / Structure and function of mammalian facilitative sugar transporters / J Biol Chem 268:19161~19164
Cope DL / 1994 / Domain assembly of the GLUT1 glucose transporter / Biochem J 300:291~294
Germann UA / 2000 / Expression of the human multidrug transporter in insect cells by a recombinant baculovirus / Biochemistry 29:2295~2303
Gorga FR / 1981 / Equilibria and kinetics of ligand binding to the human erythrocyte glucose transporter. Evidence for an alternating conformation model for transport / Biochemistry 20:5108~5113
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UCI(KEPA) : I410-ECN-0101-2013-510-002648666