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)

We developed an adenovirus vector for transduction of the human CD21 gene (Adv-CD21), the Epstein-Barr virus (EBV)-specific receptor on human B lymphocytes, to overcome the initial barrier of EBV infection in nonprimate mammalian cells. Inoculation of Adv-CD21 followed by exposure to recombinant EBV carrying a selectable marker resulted in the successful entry of EBV into three of seven nonprimate mammalian cell lines as evidenced by expression of EBV-determined nuclear antigen (EBNA). The EBV-susceptible cell lines included rat glioma-derived 9L, rat mammary carcinoma-derived c-SST-2, and canine kidney-derived MDCK. Subsequent selection culture with G418 yielded drug-resistant cell clones. In these cell clones, EBV existed as an episomal form, as evidenced through the Gardella gel technique. Among the known EBV latency-associated gene products, EBV-encoded small RNAs, EBNA1 and transcripts from the BamHI-A rightward reading frame (BARF0), and latent membrane protein 2A were expressed in all EBV-infected cell clones. The viral lytic events could be induced in these cell clones by simultaneous treatment with 12-O-tetradecanoylphorbol-13-acetate and n-butyric acid, but they were abortive, and infectious virus was not produced. These results indicate that once the initial barrier for attachment is overcome artificially, EBV can establish a stable infection in some nonprimate mammalian cells, and they raise the possibility that transgenic animals with the human CD21 gene could provide an animal model for EBV infection.
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PMID:CD21-mediated entry and stable infection by Epstein-Barr virus in canine and rat cells. 1104 19

To construct eukaryotic expression vector containing murine bcl-XL and stably express it in H18 hybridoma cells in order to enhance hybridoma cells antiapoptotic ability. PCR was used to obtain 710bp murine bcl-XL cDNA from pGEM-T-bcl-XL. Then the recombinant expression vector pEF-bcl-XL was constructed by cloning bcl-XL cDNA into eukaryotic expression vector pEF by Pst I and Xho I double digestion. After transfection into H18 hybridoma cells through lipofectamine 2000, the stable expression cell line was screened by 800mg/L G418. The expression of bcl-XL gene was detected by Western blotting. Flow cytometer was used to test the modified hybridoma cells ability to resist apoptosis induced by 0.4mmol/L Sodium Butyrate. The eukaryotic expression vector pEF-bcl-XL was successfully constructed and stably expressed in H18 hybridoma cells. Our data showed that stably transfected H18 cells expressed high levels of Bcl-XL. Under the condition of 0.4mmol/L NaBu, the production of antibody was to be significantly increased by more than 3-fold in stably transfected H18, which resulted from suppressing the NaBu-induced apoptosis and allowing stably transfected H18 cells to grow at higher viability and extend culture longevity by > 3 days. The increased culture longevity by inhibition of NaBu-induced apoptosis by inducible expression of Bcl-XL combined with the enhanced secretion of antibody by NaBu contributed to the enhancement of final antibody concentration in the stably transfected H18 cells culture. The final antibody concentration of stably transfected H18 cells in the presence of NaBu was three-fold higher than that of H18 cells culture in the absence of NaBu. Together, our results showed that butyrate is of practical interest for production of antibody. NaBu-induced apoptosis of hybridoma cells could be inhibited by inducible expression of Bcl-XL. The expression of murine bcl-XL gene in hyridoma cells and the increasing antiapoptosis ability of hybridoma cells are of significance in further use of hybridoma cells in high density large scale cell culture.
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PMID:[Modification of the antiapoptotic ability of H18 hybridoma cells]. 1597 83