Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: DrugBank:EXPT01586 (G418)
 2,237 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A retroviral vector carrying the E6 and E7 genes of HPV type 18 was transfected into a packaging cell line, Ampho psi 2. Thirteen recombinant viruses carrying the E6 and E7 genes were obtained. The titers of these recombinant viruses were estimated by infecting BALB/c3T3 cells and then counting the number of G418r colonies. Presence of HPV E6/E7 genes was confirmed by the PCR method and sequence-specific primers. The expression of E7 gene was examined by RT-PCR method. Results showed that the titers were ranged between 0.2 and 1.2 x 10(3) CFU/ml and the E7 transcripts were detected in all 13 cell clones. These E6 and E7-containing cell clones were able to grow in soft agar, indicating the E6/E7 delivered by the recombinant retroviruses retained their transformation function. These recombinant viruses were then used to infect human NPC cell lines, NPC-TW076 and -TW039 and cell clones resistant to G418 were obtained. Using Western blot analysis and HPV type 18 E6-specific monoclonal antibody, HPV-CIP5, these cells were shown to contain a protein with a molecular mass of 18 kDa. Our data indicated that the HPV E6/E7-containing recombinant retroviruses were capable of infecting human cell lines. The potential of using these recombinant retroviruses to immortalize human primary epithelial cells was discussed.
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PMID:Transfer of the E6 and E7 genes of human papillomavirus type 18 into human epithelial cells via recombinant retrovirus infection. 1059 18

Human neural progenitor cells (hNPCs) are a promising source to treat various neurodegenerative diseases. Potential applications are to use such cells for reprogramming to induce pluripotent stem cells or for secretion of proteins into the brain. These applications usually involve expression of heterologously expressed genes which is difficult to achieve in hNPCs. We tested several protocols for non-viral gene transfer and different promoters. Nucleofection and the cytomegalovirus enhancer/chicken beta-actin promoter allowed expression of foreign genes in hNPCs for up to 6 months. Treatment with the antibiotic G418 enabled us to select stably transfected cells which were subcloned and continued to express the NPC marker nestin. Differentiation of stably nucleofected hNPCs revealed that multipotency was maintained following long-term expansion of subcloned hNPCs. After differentiation for 3 weeks in vitro or in vivo following striatal transplantations transfected hNPCs expressed voltage-gated sodium channels suggesting the development of functional properties during neuronal maturation. In conclusion, stably nucleofected hNPCs can be isolated, subcloned, and expanded for up to 6 months without loss of their differentiation potential. These data provide a basis for future studies using hNPCs to investigate the neuronal differentiation in vivo after transplantation, the feasibility as a vector for gene (protein) therapy, and the induction of pluripotent stem cells.
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PMID:Non-viral gene transfer by nucleofection allows stable gene expression in human neural progenitor cells. 1905 35