Expression and purification of recombinant nattokinase in Spodoptera frugiperda cells.
A recombinant baculovirus, rv-egfp-NK, containing a reporter gene encoding the enhanced green fluorescent protein (EGFP), was used to express nattokinase (NK), a fibrinolytic enzyme, in Spodoptera frugiperda (SF-9) cells. The recombinant protein also included a histidine tag for purification using Ni(2+) resins. The recombinant NK, approximately 30 kDa, retained fibrinolytic activity (60 U/ml). The integration of the EGFP expression cassette in the Bac-to-Bac system is thus an effective method for the expression and purification of recombinant NK protein in Spodoptera frugiperda insect cells.
Li
Xiaoxiang
X
College of Life Sciences, Hubei University, Wuhan, Hubei, 430062, China.
Wang
Xiaoli
X
Xiong
Shaoling
S
Zhang
Jing
J
Cai
Litao
L
Yang
Yanyan
Y
D000818:Animals; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D015971:Gene Expression Regulation, Enzymologic; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D018411:Spodoptera; D013381:Subtilisins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D013381:Subtilisins; C053771:nattokinase