Monitoring of the effects of transfection with baculovirus on Sf9 cell line and expression of human dipeptidyl peptidase IV

被引:9
作者
Ustun-Aytekin, Ozlem [1 ,2 ,3 ]
Gurhan, Ismet Deliloglu [2 ]
Ohura, Kayoko [3 ]
Imai, Teruko [3 ]
Ongen, Gaye [2 ]
机构
[1] Pamukkale Univ, Fac Engn, Dept Food Engn, Kinikli, Denizli, Turkey
[2] Ege Univ, Fac Engn, Bioengn Dept, TR-35100 Izmir, Turkey
[3] Kumamoto Univ, Grad Sch Pharmaceut Sci, Kumamoto, Japan
关键词
Spodoptera frugiperda (Sf9); Dipeptidyl peptidase IV (DPPIV); Baculovirus; Transfection; TRANSIENT TRANSFECTION; AMINOPEPTIDASE; SERUM; PURIFICATION; FRUGIPERDA; AUTISM; GENE;
D O I
10.1007/s10616-013-9549-3
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Human dipeptidylpeptidase IV (hDPPIV) is an enzyme that is in hydrolase class and has various roles in different parts of human body. Its deficiency may cause some disorders in the gastrointestinal, neurologic, endocrinological and immunological systems of humans. In the present study, hDPPIV enzyme was expressed on Spodoptera frugiperda (Sf9) cell lines as a host cell, and the expression of hDPPIV was obtained by a baculoviral expression system. The enzyme production, optimum multiplicity of infection, optimum transfection time, infected and uninfected cell size and cell behavior during transfection were also determined. For maximum hDPPIV (269 mU mL(-1)) enzyme, optimum multiplicity of infection (MOI) and time were 0.1 and 72 h, respectively. The size of infected cells increased significantly (P < 0.001) after 24 h post infection. The results indicated that Sf9 cell line was applicable to the large scale for hDPPIV expression by using optimized parameters (infection time and MOI) because of its high productivity (4.03 mU m L-1 h(-1)).
引用
收藏
页码:159 / 168
页数:10
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