Purification and characterization of human immunodeficiency virus type 1 nef gene product expressed by a recombinant baculovirus.
We have constructed the recombinant baculovirus which expresses the human immunodeficiency virus type 1 negative factor (nef) gene. Spodoptera frugiperda cells infected with the recombinant virus produced a 27-kDa protein which reacted with rabbit antisera raised against a carboxy-terminal synthetic peptide of the Nef protein by immunoblot analysis. Labeling experiment showed that the recombinant Nef protein was myristoylated. The recombinant Nef protein was purified to near homogeneity by DEAE-Sephacel, phenyl-Sepharose 4B, blue-Sepharose, and Sephadex G-150 column chromatography. No detectable GTP binding activity was observed in the purified recombinant Nef product.
Matsuura
Y
Y
Department of Veterinary Science, National Institute of Health, Tokyo, Japan.
Maekawa
M
M
Hattori
S
S
Ikegami
N
N
Hayashi
A
A
Yamazaki
S
S
Morita
C
C
Takebe
Y
Y
D000818:Animals; D016367:Baculoviridae; D003001:Cloning, Molecular; D019204:GTP-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism; D015693:Gene Products, nef / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D015483:HIV Antibodies / Q000276:immunology*; D015488:HIV Antigens / Q000276:immunology; D015658:HIV Infections / Q000276:immunology; D006679:HIV Seropositivity / Q000276:immunology; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D009036:Moths / Q000378:metabolism; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D054311:nef Gene Products, Human Immunodeficiency Virus
D015693:Gene Products, nef; D015483:HIV Antibodies; D015488:HIV Antigens; D009227:Myristic Acids; D011994:Recombinant Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; D019814:Myristic Acid; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins