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 have developed a gene trap approach to select specific cytokine receptor/ligand responsive genes in the cell line TF-1. This cell line exhibits a dependency on granulocyte-macrophage colony-stimulating factor (GM-CSF) or interleukin-3 (IL-3) and responds to interleukin-5 (IL-5). In an attempt to detect genes modulated by one of these factors, cells were infected with the Rosabetageo retrovirus in the presence of GM-CSF, IL-3, or IL-5 and clones were selected for retroviral integration on the basis of G418 resistance. Housekeeping and cytokine-regulated trapped genes were then differentiated on the basis of G418 resistance versus sensitivity in the presence of the different cytokines. To determine the reliability of this screen, DNA sequences upstream of the proviral integration site were identified by 5' rapid amplification of DNA ends polymerase chain reaction (RACE PCR) from selected GM-CSF-treated and -infected clones. Comparison of the sequences with those in the Genbank database revealed that 2 sequences correspond to known genes: NACA and RBM3. NACA was recently defined as a coactivator of c-jun-mediated transcription factors in osteoblasts, and RBM3 as a protein from the heterogeneous nuclear ribonucleoprotein family. Data from transcriptional analysis of these 2 genes in TF-1 cells showed a specific up-regulation by GM-CSF. Both transcripts were also found to be up-regulated in purified CD34(+) cells, suggesting their involvement in proliferative processes during hematopoiesis. Interestingly, down-regulation was observed during monocytic differentiation of TF-1 cells, suggesting their extinction could contribute to monocytic lineage development. This study demonstrates that this gene trap approach is a useful method for identifying novel, specific cytokine-responsive genes that are involved in the regulation of hematopoiesis. (Blood. 2000;95:3750-3757)
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PMID:Capture of cytokine-responsive genes (NACA and RBM3) using a gene trap approach. 1084 6

In order to look for the tumor-associated genes from human multiple myeloma (MM), a cDNA library of human multiple myeloma cell line ARH-77 was constructed with eukaryote expression vector pcDNA3.1(+). The length of inserted fragments in library was 1.2 kb in average. All clones in cDNA library were transferred in situ to nylon membrane, which was divided into eight equal parts (A-H) and cultured in LB medium to set up gene pools. The plasmids in cDNA library and in gene pools were extracted and NIH/3T3 cells were transfected respectively. By G418 screening and colonies counting, gene pool A was chosen for the second cycle transfection. After several cycles, a clone, A62-17, was obtained, which had significant transforming ability. The length of this clone was 993 bp. The RACE technique was used for rapid amplification of A62-17 5'-end. The full length of this sequence has 1300 bp and was named as hMMTAG2 gene. hMMTAG2 consists of 8 exons and codes for a polypeptide of 263 amino acids (the accession number in GenBank: AY137773). It was located at chromosome 1q42.13. hMMTAG2 had same transforming activities in NIH/3T3 cells as the clone A62-17, and the number of transformant foci was 6 folds more than the blank vector pcDNA3.1(+). The analysis of bioinformatics revealed that hMMTAG2 had many phosphorylation sites for several protein kinases, N-myristoylation sites and nuclear localization signals, so it may be a signal molecule in the nucleus.
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PMID:Cloning and sequence analysis of tumor-associated gene hMMTAG2 from human multiple myeloma cell line ARH-77. 1254 21

Porcine CFL2b gene play an important role in the muscle development and myofibrillar formation in pig. To explore whether CFL2b expression affects muscle fiber trait, the porcine CFL2b full-length cDNA was amplified using homology based cDNA cloning and SMART RACE. Then the full length cDNA of porcine CFL2b was inserted into pEGFP-N1 and transfected into C2C12 cells. The cells stably expressing CFL2b were selected by G418. We examined the expression of MyHC 2x, MyHC 2b and MyHC1/slow in C2C12 cells stably expressing CFL2b. The results showed that the level of MyHC 2x and MyHC 2b mRNA were dramatically increased compared with control cells, while the level of MyHC1/slow mRNA is not changed. To identify the transcription events of CFL2b, the porcine CFL2b mRNA was detected by Northern blotting, two transcripts, long transcript (3,012 bp) and short transcript (1,466 bp) were found in porcine skeletal muscles. The nucleotide sequence of CFL2b shares 88.1 and 74.9% homology with the CFL2b gene in human and mouse. The deduced amino acid sequence of CFL2b (166 amino acids) in pig shares 100, 99.1% identity with the CFL2b in human and mouse, respectively. Taken together, our research revealed that porcine CFL2b may be involved in the regulation muscle fiber trait by affecting the expression of MyHC.
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PMID:The full length cloning of a novel porcine gene CFL2b and its influence on the MyHC expression. 1912 43