Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Artificial Gene Fusion”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 19 recordsLinked to original sources
PubMed2001

Human monocyte x mouse melanoma fusion hybrids express human gene.

Artificial fusion of human monocyte with Cloudman S91 mouse melanoma cells resulted in hybrids that showed increased motility in vitro, enhanced metastatic potential in vivo, and also tended to be super melanotic (Rachkovsky et al., Clin. Exp. Metastasis 16 (1998) 299). However, no gene derived from monocytes has been shown to be expressed in these hybrids until now. Similar observations have also been noted in hybrids originating from mouse macrophage and mouse melanoma cells. Having the advantage of species differences in mouse x human hybrids, we are able, this time, to show by RT-PCR that some genes specific to the human genome are expressed in these hybrids, indicating that not only is the genomic DNA from parental monocytes integrated in the hybrids but also some genes are being expressed. This observation may lead us to find contributory genes from monocyte and/or macrophage that are responsible for modulating the genotypes and hence the phenotypes in the hybrids.

Animals↗
PubMed1987

Characterization of an artificial bifunctional enzyme, beta-galactosidase/galactokinase, prepared by gene fusion.

An artificial bifunctional enzyme, beta-galactosidase/galactokinase, has been prepared by gene fusion. The hybrid protein catalyzes the hydrolysis of lactose followed by phosphorylation of galactose. The protein has been purified using DEAE-Sephacel chromatography and gel filtration on Sephacryl S-400 Superfine. The configuration of the hybrid protein is mainly tetrameric but also higher aggregates can be detected. The monomer Mr is 160,000 as judged from sodium dodecyl sulfate/polyacrylamide gel electrophoresis and the native Mr has been calculated to be 600,000-650,000 from gel filtration experiments. beta-Galactosidase/galactokinase has different thermostability curves, pH/activity profiles and Km values as compared with the native enzymes. By using a third enzyme, galactose dehydrogenase, which competes with galactokinase for the galactose formed by beta-galactosidase, substrate channeling can be detected. This proximity effect becomes even more pronounced in an assay mixture containing poly(ethylene glycol).

Chromatography, Gel↗
PubMed1992

Dual initiation sites of protein synthesis on foot-and-mouth disease virus RNA are selected following internal entry and scanning of ribosomes in vivo.

The initiation of protein synthesis on foot-and-mouth disease virus RNA occurs at two sites separated by 84 nucleotides. Immediately upstream from the first of these sites is the internal ribosome entry site (IRES), which directs the translation of this RNA to be cap-independent. The utilization of these two initiation sites has been examined using artificial fusion genes in vivo under a variety of conditions. Additional in-frame AUG codons have been introduced between these two authentic start sites to determine the mechanism by which ribosomes recognize the second start site. The results indicate that following internal entry of ribosomes on the 5' side of the first initiation codon, many fail to initiate protein synthesis at this position and scan along the RNA to the second initiation site. In the presence or absence of the IRES both initiation sites are efficiently used but the utilization of the two sites is slightly biased towards the second initiation site by the IRES. Furthermore, in the presence of the IRES, protein synthesis initiates at both sites independently of the activity of the cap-binding complex.

Aphthovirus↗
PubMed2004

A novel FISH assay for SS18-SSX fusion type in synovial sarcoma.

Synovial sarcoma is a morphologically, clinically and genetically distinct entity that accounts for 5-10% of all soft tissue sarcomas. The t(X;18)(p11.2;q11.2) is the cytogenetic hallmark of synovial sarcoma and is present in more than 90% of the cases. It produces three types of fusion gene formed in part by SS18 from chromosome 18 and by SSX1, SSX2 or, rarely, SSX4 from the X chromosome. The SS18-SSX fusions do not seem to occur in other tumor types, and it has been shown that in synovial sarcoma a clear correlation exists between the type of fusion gene and histologic subtype and, more importantly, clinical outcome. Previous analyses regarding the type of fusion genes have been based on PCR amplification of the fusion transcript, requiring access to good-quality RNA. In order to obtain an alternative tool to diagnose and follow this malignancy, we developed a fluorescence in situ hybridization (FISH) assay that could distinguish between the two most common fusion genes, that is, SS18-SSX1 and SS18-SSX2. The specificity of the selected bacterial artificial chromosome clones used in the detection of these fusion genes, as well as the sensitivity of the analysis in metaphase and interphase cells, was examined in a series of 28 synovial sarcoma samples with known fusion gene status. In all samples, the type of fusion was correctly identified by FISH. Thus, the assay described here should be useful for clarifying unresolved chromosome markers and for identifying fusion gene status in samples from which RNA of sufficient quality for PCR could not be extracted.

Antigens, Neoplasm↗
PubMed2000

Gene fusions.

Explore the source record for details and available documents.

Artificial Gene Fusion↗
PubMed2002

Design of endonuclease restriction sites into primers for PCR cloning.

I present a software system PCRCLNG that facilitates the design of endonuclease restriction sites into the 5'-end of PCR primers. The product amplified using these primers can be directly cloned into vectors. The program estimates the annealing temperature for each primer and selects the primer pairs with comparable annealing temperature. Finally the software determines whether the PCR product can be cloned into the vector to generate in-frame gene fusion.

Algorithms↗
PubMed1999

Insertional gene fusion technology.

The classical 'end to end' gene fusion technique has widely been used for monitoring gene expression, biological screening and purification of recombinant proteins. Recent progress with the 'insertional' gene fusion approach, on the other hand, has demonstrated that this technique can be utilized for membrane protein topology analysis, display of randomized protein libraries and design of biosensor proteins. In this review, we describe examples of insertional gene fusion and compare the old and new gene fusion techniques.

Artificial Gene Fusion↗
PubMed2001

A ParE-ParC fusion protein is a functional topoisomerase.

Type II topoisomerases are responsible for DNA unlinking during DNA replication and chromosome segregation. Although eukaryotic enzymes are homodimers and prokaryotic enzymes are heterotetramers, both prokaryotic and eukaryotic type II topoisomerases belong to a single protein family. The amino- and carboxyl-terminal domains of eukaryotic enzymes are homologous to the ATP-binding and catalytic subunits of prokaryotic enzymes, respectively. Topoisomerase IV, a prokaryotic type II topoisomerase, consists of the ATP-binding subunit, ParE, and the catalytic subunit, ParC. We have joined the coding regions of parE and parC in frame and constructed a fusion protein of the two subunits of topoisomerase IV. This fusion protein, ParEC, can catalyze both decatenation and relaxation reactions. The ParEC protein is also capable of decatenating replicating daughter DNA molecules during oriC DNA replication in vitro. Furthermore, the fusion gene, parEC, complements the temperature-sensitive growth of both parC and parE strains, indicating that the ParEC protein can substitute for topoisomerase IV in vivo. These results demonstrate that a fusion protein of the two subunits of topoisomerase IV is a functional topoisomerase. Thus, a heterotetrameric type II topoisomerase can be converted into a homodimeric type II topoisomerase by gene fusion.

Artificial Gene Fusion↗
PubMed1991

Multienzyme systems obtained by gene fusion.

The preparation of artificial bi- and polyfunctional enzymes by gene fusion appears to have great potential in enzyme technology. Their purification is facilitated compared with that of their naturally occurring counterparts and they exhibit favourable enzyme kinetics. Applications can be found in biochemical analysis, enzyme process technology and metabolic engineering.

Cloning, Molecular↗
PubMed2001

A multiplex real-time pcr assay for the detection of gene fusions observed in solid tumors.

SUMMARY: Specific gene fusions observed in solid tumors are extremely useful diagnostic markers. We report the development of a method based on real-time PCR which enables the detection upon identical PCR conditions of the different fusions specifically observed in Ewing tumors (ET), alveolar rhabdomyosarcoma (ARMS), synovial sarcoma (SS), small round cell desmoplastic tumors (SRCDT), extraskeletal myxoid chondrosarcoma, malignant melanoma of soft parts, congenital fibrosarcoma, and anaplastic large cell lymphoma. A simple assay, based on multiplexing of primers and probes, is described for the routine genetic diagnosis of small round cell tumors of children. It enables the detection of the five EWS-ETS, the two PAX-FKHR, the three SYT-SSX, and the EWS-WT1 fusions of ET, ARMS, SS, and SRCDT, respectively. The sensitivity of this test is high enough to detect all fusions, including the large EWS-FLI-1 transcripts, with the equivalent of 100 tumor cells as a starting material. This multiplex fluorescent analysis of chromosome translocations (MFACT) was validated in comparison with conventional RT-PCR on a series of 79 tumors. A major advantage of this method is that it completely abolishes the manipulation of PCR-products. It, therefore, considerably lowers the risk of cross-contamination linked to carry-over of RT-PCR products. It also constitutes an important step toward the complete automation of the detection of cancer-specific gene fusions.

Artificial Gene Fusion↗
PubMed1988

Studies on the import into mitochondria of yeast ATP synthase subunits 8 and 9 encoded by artificial nuclear genes.

Direct fusions have been constructed between each of subunits 8 and 9 from mitochondrial ATPase of Saccharomyces cerevisiae, proteins normally encoded inside mitochondria, and the cleavable N-terminal transit peptide from the nuclearly encoded precursor to subunit 9 of Neurospora crassa mitochondrial ATPase. The subunit 8 construct was imported efficiently into isolated yeast mitochondria and was processed at or very near the fusion point. When expressed in vivo from its artificial nuclear gene, this cytoplasmically synthesized form of subunit 8 restored the growth defects of aap 1 mutants unable to produce subunit 8 inside the mitochondria. The subunit 9 construct was, however, unable to be imported into isolated mitochondria and could not, following nuclear expression in vivo, complement growth defects in mitochondrial oli 1 mutants. This behaviour is contrasted with the previously demonstrated import competence of another yeast subunit 9 fusion, bearing the first five residues of mature N. crassa subunit 9 interposed between its own transit peptide and the yeast subunit 9 moiety.

Amino Acid Sequence↗
PubMed2002

Preparation of luciferase-bacterial magnetic particle complex by artificial integration of MagA-luciferase fusion protein into the bacterial magnetic particle membrane.

MagA is an iron-translocating protein found in the membranes of magnetic bacterium. Luciferase-bacterial magnetic particle (BMP) complexes were prepared by artificially inserting MagA-luciferase fusion proteins into the membranes of BMPs from Magnetospirillum magneticum strain AMB-1. Fusion proteins were from recombinant Escherichia coli membranes. MagA-Luc fusion proteins were integrated by sonication in vitro. Successful integration of fusion proteins was confirmed by luciferase luminescence on BMPs. Maximum luminescence was obtained after sonication for 3 min with a solution containing 300 mM NaCl, and is 18 times higher compared with recombinant Luc-BMPs generated using previously reported gene fusion techniques.

Artificial Gene Fusion↗