1. Cloning and transmembrane glycoprotein expression of the retrovirus HTLV-1 in mammals' cells
- Author
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Luciene Medeiros, Maristela Delgado Orellana, Dimas Tadeu Covas, Osvaldo Massaiti Takayanagui, Patricia Vianna Bonini Palma, Flora Cristina Lobo Penteado, Aparecida Maria Fontes, Universidade de São Paulo (USP), and Universidade Estadual Paulista (Unesp)
- Subjects
glycoprotein ,molecular cloning ,polymerase chain reaction ,animal cell ,immunogenicity ,law.invention ,Retrovirus ,Viral Envelope Proteins ,law ,Tropical spastic paraparesis ,spinal cord disease ,membrane protein ,Mammals ,Human T-lymphotropic virus 1 ,Membrane Glycoproteins ,biology ,Structural gene ,Gene Expression Regulation, Developmental ,cell line ,gene control ,cytometry ,Flow Cytometry ,tropical disease ,Glicoproteína transmembrana ,Infectious Diseases ,Mammalia ,virus gene ,Recombinant DNA ,Microbiology (medical) ,spastic paresis ,Cloning, Organism ,structural gene ,Transmembrane glycoprotein ,Heterologous ,Molecular cloning ,envelope gene ,medicine ,Animals ,Animalia ,Cell Lineage ,Clonagem ,nonhuman ,DNA fragment ,T cell leukemia ,HEK 293 cells ,biology.organism_classification ,medicine.disease ,Virology ,Molecular biology ,HTLV-1 ,protein blood level ,Protein expression ,Parasitology ,genetic transfection ,Human T cell leukemia virus 1 ,Expressão de proteínas ,recombinant protein ,Cloning - Abstract
Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:21:49Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:38:09Z : No. of bitstreams: 1 2-s2.0-33646676282.pdf: 608428 bytes, checksum: a93d671ba39fdc01a516243c0dbcc135 (MD5) Made available in DSpace on 2014-05-27T11:21:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-03-01 The retrovirus HTLV-1 is the etiological agent of the adult T-cell leukemia and HTLV-1 associated myelopathy/tropical spastic paraparesis. The proviral genome has 9,032 base pairs, showing regulatory and structural genes. The env gene encodes for the transmembrane glycoprotein gp 21. The development of methodologies for heterologous protein expression, as well as the acquisition of a cellular line that constituently expresses the recombinant, were the main goals of this work. The DNA fragment that encodes for gp 21 was amplified by nested-PCR and cloned into a pCR2.1-TOPO vector. After which, a sub-cloning was realized using the expressing vector pcDNA3.1+. The transfection of mammalian cells HEK 293 was performed transitorily and permanently. Production of the recombinant gp 21 was confirmed by flux cytometry experiments and the cell line producing protein will be used in immunogenicity assays. Laboratório de Pesquisa Centro Regional de Hemoterapia Universidade de São Paulo, Ribeirão Preto, SP Departamento de Análises Clínicas Faculdade de Ciências Farmacêuticas Júlio de Mesquita Filho Universidade Estadual Paulista, Araraquara, SP Departamento de Neurologia Faculdade de Medicina de Ribeirão Preto Universidade de São Paulo, Ribeirão Preto, SP Departamento de Clinica Medica Faculdade de Medicina de Ribeirão Preto Universidade de São Paulo, Ribeirão Preto, SP Centro Regional de Hemoterapia HCFMRP/Hemocentro de Ribeirão Preto. R. Tenente, Catão Roxo 2501, 14051-140 Ribeirão Preto, SP Departamento de Análises Clínicas Faculdade de Ciências Farmacêuticas Júlio de Mesquita Filho Universidade Estadual Paulista, Araraquara, SP
- Published
- 2006