Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.21.1 (DNase)
 7,655 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Recombinant adeno-associated virus 2 (AAV) virions were constructed that contained the genomic copy of a normal human beta-globin gene marked with a 4-bp Clal linker, and the herpesvirus thymidine kinase (TK) promoter-driven bacterial gene for resistance to neomycin (v beta m-globin), as well as those containing the DNase l-hypersensitive site 2 (HS-2) from the locus control region (LCR) of the human beta-globin gene cluster (vHS2-beta m-globin). These recombinant virions were used to infect a human erythroleukemia cell line which normally does not express the beta-globin gene (K562), or a human nasopharyngeal carcinoma cell line (KB). Cell populations resistant to G418, a neomycin analogue, were obtained following infections with the recombinant virions, indicating high-efficiency transduction of the chimeric gene as well as functional activity of the transduced neo gene in both cell types. Southern blot analysis using a human beta-globin DNA probe substantiated stable integration of the exogenous beta-globin allele in these cells. There was no expression of the transduced beta-globin gene in K562 or KB cells infected with the v beta m-globin virus. High-level expression of the transduced beta-globin gene occurred only in the vHS2-beta m-globin virus-infected K562 cells, but not in KB cells, as determined by Northern blot as well as RNase protection analyses. Expression of the human beta-globin protein could also be detected in approximately 10-20% of the vHS2-beta m-globin virus-infected K562 cells. These studies suggest that the AAV-based vector system may prove useful for high-efficiency globin gene transfer in human hematopoietic cells.
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PMID:Adeno-associated virus 2-mediated transduction and erythroid cell-specific expression of a human beta-globin gene. 864 53

This study was aimed to construct the CD14 eukaryotic expression vector, establish the transgeneic CD14 positive cell line in order to facilitate the establishment of a mouse model of antibody targeting therapy for human acute monocytic leukemia (AML-M(5)). Total RNA extracted from peripheral blood mononuclear cells was treated with RNAase-free DNAase, the human CD14 gene was cloned and sequenced through the RT-PCR and T-A clone techniques. Eukaryotic expressional vector pcDNA3.1(+)/CD14 was constructed by cleaving with double restriction endonuleases and ligating with T4 ligase. A murine melanoma cell line B16 was transfected with the pcDNA3.1(+)/CD14 recombinant with Superfect transfection reagent. Positive clones were selected by G418 and the expression of human CD14 on the transfectant was confirmed by flow cytometry (FCM). The results indicated that the sequence of the human CD14 cDNA cloned was exact to be same as the one from GenBank database. The recombinant pcDNA3.1(+)/CD14 was identified with double-enzyme cleaving. The expression of the human CD14 on the transfectant (B16/CD14) was confirmed by FCM. In conclusion, the murine cell line B16/CD14 fransfected with human CD14 gene has been established which can be used for the study of human AML-M(5) antibody targeting therapy with mouse model.
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PMID:[Establishment of murine cell line transfected with human CD14 gene]. 1663 22