RESUMO
A plasmid named pSH-G was constructed with the rabies-virus G-gene insert. This plasmid was transfected into eukaryotic BHK-21 cells and its stability tested. The presence of the pSH-G plasmid was confirmed by means of polymerase chain reaction (PCR) after each of ten cell passages, and the results were positive. The stable BHK-21/pSH-G+ clone obtained can be used in the study of rabies as well as in the production of vaccines.
Assuntos
Glicoproteínas/genética , Plasmídeos , Proteínas Recombinantes de Fusão/genética , Vírus da Raiva , TransfecçãoRESUMO
Treatment of cancer using gene therapy is based on adding a property to the cell leading to its elimination. One possibility is the use of suicide genes that code for enzymes that transform a pro-drug into a cytotoxic product. The most extensively used is the herpes simplex virus thymidine kinase (TK) gene, followed by administration of the antiviral drug ganciclovir (GCV). The choice of the promoter to drive the transcription of a transgene is one of the determinants of a given transfer vector usefulness, as different promoters show different efficiencies depending on the target cell type. In the experiments presented here, we report the construction of a recombinant adenovirus carrying TK gene (Ad-TK) driven by three strong promoters (P CMV IE, SV40 and EN1) and its effectiveness in two cell types. Human HeLa and mouse CCR2 tumor cells were transduced with Ad-TK and efficiently killed after addition of GCV. We could detect two sizes of transcripts of TK gene, one derived from the close together P CMV IE/SV40 promoters and the other from the 1.5 Kb downstream EN1 promoter. The relative amounts of these transcripts were different in each cell type thus indicating a higher flexibility of this system
Assuntos
Humanos , Animais , Camundongos , Adenoviridae , Antivirais , Ganciclovir , Terapia Genética , Timidina Quinase , Genes Virais , Vetores Genéticos , Células HeLa , Regiões Promotoras Genéticas , Células Tumorais CultivadasRESUMO
Malignant transformation is accompanied by changes in cell-matrix interations. Upon transfection with EJ-ras oncogene, transformed fibroblasts, acquired a migratory phenotype towards laminin-1. The increase in integrin expression was responsible for the migratory activity of transformed fibroblasts. In addition alpha(6)beta(1) integrins, both galectin-3 and an unidentified laminin-binding polypeptide had their expression pattern altered upon transformation. Here, we review these two classes of laminin-binding proteins and their possible roles in cell-laminin interactions.