Expression of the HadV-41 short fiber C-terminal knob domain in baculovirus system (2001)
- Authors:
- USP affiliated authors: SILVA, JOÃO SANTANA DA - FMRP ; MEHNERT, DOLORES URSULA - ICB ; HARSI, CHARLOTTE MARIANNA - ICB
- Unidades: FMRP; ICB
- Assunto: MICROBIOLOGIA
- Language: Inglês
- Abstract: Human adenovirus of Subgenus F, serotypes 40 and 41, cause infantile gastroenteritis. Structural features uniques of these viruses concern the presence of two different fibers in the viral particle: a long and a short, placed one per penton-base. The long fiber, probably, undergoes the viral adsorption to the host cell by interaction with the receptor CAR. The function of the short fiber is not known. The aim of this project is obtain the Ad41 short fiber knob domain purified, to use it as a ligand on virus-host cell interaction studies. The viral DNA sequence corresponding to the C-terminal domain of the short fiber of the HAdV-41(Tak) was amplified by PCR and cloned into pFastBacDual plasmid (Gibco-BRL). E. coli DH10Bac competent cells, which contain a modified baculovirus genome (Bacmid-Gibco-BRL) were transformed with the donor plasmid. The cells were plated on selective medium and the colonies were screened for their lacZ phenotype (white/blue). High molecular weight mini-prep DNA was prepared from selected E.coli clones. Verification of the insertion of the Ad41-short fiber sequence into the Bacmid DNA was done by PCR with the pUC/M13 (Gibco-BRL) amplification primers. The recombinant Bacmid DNA was then used to transfect Sf21 insect cells. The recombinant baculovirus was recovered from the cultures. Expression trials were done on Sf21, HighFive and Sf+ insect cell lines. The expressed protein was detected by Western-blot assays using an anti-Ad41short-fiber rabbit serum. Expression kinetics assays showed best results on Sf21 cells, collected three days post-infection
- Imprenta:
- Publisher: Comissão de Cultura e Extensão Universitária do ICB/USP
- Publisher place: São Paulo
- Date published: 2001
- Source:
- Título do periódico: Resumos
- Conference titles: Congresso do Instituto de Ciências Biomédicas
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ABNT
FAVIER, A L et al. Expression of the HadV-41 short fiber C-terminal knob domain in baculovirus system. 2001, Anais.. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP, 2001. . Acesso em: 24 abr. 2024. -
APA
Favier, A. L., Yeda, F. P., Silva, J. S. da, Mehnert, D. U., Gout, E., van Raaji, M. J., et al. (2001). Expression of the HadV-41 short fiber C-terminal knob domain in baculovirus system. In Resumos. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP. -
NLM
Favier AL, Yeda FP, Silva JS da, Mehnert DU, Gout E, van Raaji MJ, Kidd A, Chroboczek J, Hársi CM. Expression of the HadV-41 short fiber C-terminal knob domain in baculovirus system. Resumos. 2001 ;[citado 2024 abr. 24 ] -
Vancouver
Favier AL, Yeda FP, Silva JS da, Mehnert DU, Gout E, van Raaji MJ, Kidd A, Chroboczek J, Hársi CM. Expression of the HadV-41 short fiber C-terminal knob domain in baculovirus system. Resumos. 2001 ;[citado 2024 abr. 24 ] - Expression of the c-terminal domain (KNOB) of HADV-41 short fiber in baculovirus system
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