Cloning and transmembrane glycoprotein expression of the retrovirus HTLV-1 in mammals' cells

Revista da Sociedade Brasileira de Medicina Tropical

Orellana, Maristela Delgado¹  Universidade Estadual Paulista, Araraquara, Brasil¹  Covas, Dimas Tadeu¹  Penteado, Flora Cristina Lobo¹  Universidade de São Paulo, Ribeirão Preto, Brasil¹  Palma, Patricia¹  Medeiros, Luciene¹  Takayanagui, Osvaldo Massaiti¹  Fontes, Aparecida Maria¹ 

关键词: Transmembrane glycoprotein;    HTLV-1;    Cloning;    Protein expression;   

DOI  :  10.1590/S0037-86822006000200007

学科分类：农业科学（综合）

来源: Sociedade Brasileira de Medicina Tropical

【 摘 要 】

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.

【 授权许可】

Unknown