A new Trichoplusia ni cell line for membrane protein expression using a baculovirus expression vector system.
A new cell line, MSU-TnT4 (TnT4), was established from Trichoplusia ni embryos for use with baculovirus expression vectors and evaluated for its potential for membrane protein production. To evaluate membrane protein synthesis, recombinant baculoviruses were constructed to express the human neurotensin receptor 1 as an enhanced green fluorescent protein (GFP) fusion. TnT4 cells had a doubling time of 21 h and expressed the membrane-GFP fusion protein at approximately twice the level as Sf21 cells from the p10 promoter, as evaluated by GFP intensity. Expression of secreted alkaline phosphatase (SEAP) was similar to that of Sf21 cells. Expression of membrane-GFP fusion proteins in recombinant baculoviruses provides a rapid method for evaluating the potential of new cell lines for the production of membrane proteins using a baculovirus expression vector system (BEVS).
Zhang
Fengrui
F
Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, 48825, USA.
Manzan
Maria Alejandra
MA
Peplinski
Heather M
HM
Thiem
Suzanne M
SM
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D049109:Cell Proliferation; D004587:Electrophoresis, Agar Gel; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics*; D006023:Glycoproteins / Q000096:biosynthesis; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D018411:Spodoptera / Q000166:cytology*; D013347:Subcellular Fractions / Q000378:metabolism
D006023:Glycoproteins; D008565:Membrane Proteins; D049452:Green Fluorescent Proteins; D000469:Alkaline Phosphatase