Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.30.2 (endonuclease)
 18,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A beta-actin gene of carp (Cyprinus carpio) was isolated from a genomic EMBL3 library. The nucleotide sequence of the gene indicates six exons spanning 3.6 kb. Southern blot hybridization of restriction endonuclease digests of carp genomic DNA indicate that there are two copies of the beta-actin isotype and several other species of actin genes. The transcriptional start site is 85 bp and 24 bp downstream respectively from consensus CCAAT and TATA promoter elements. The organization of the carp beta-actin gene is identical to that of chicken, human, and rat genes in terms of size, exon/intron locations and junctions and in having a translationally silent first exon. The fish gene is 90% and 99% conserved at the nucleotide and amino acid levels, respectively, with land vertebrate beta-actin genes. Northern blot analysis of beta-actin gene expression indicated that the gene is highly expressed in brain, less so in muscle, and much less so in liver cells. The putative beta-actin proximal promoter of carp, identified by the conservation of known actin regulatory sequences, is transcriptionally active in both mammalian and piscine cells.
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PMID:Isolation and characterization of beta-actin gene of carp (Cyprinus carpio). 213 83

Many genes have been transferred into fish for scientific and aquacultural purposes. We have been developing expression vectors containing regulatory sequences from the carp beta-actin gene enhancer/promoter for expression of genes or cDNAs in transgenic fish. Expression from these vectors varies over a 20-fold range in zebrafish, beginning within 12 hours of fertilization and continuing for at least two weeks. Expression can be found in nearly all tissues. The vectors have the following characteristics: (1) they contain either unique or polycloning restriction endonuclease sites for insertion of any gene or cDNA, and (2) the piscine sequences are flanked by restriction sites for easy removal of plasmid, or nonfish, sequences. We have tested the ability of special sequences, border elements, from other animals to confer position-independent expression of transgenes or enhance integration of transgenic constructs into fish chromosomes. Early results indicate that these elements do not act as enhancers and do not improve integration frequencies. However, both avian and insect border elements are able to confer position-independent expression as judged from expression of CAT genes in F1 generation fish.
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PMID:Development of position-independent expression vectors and their transfer into transgenic fish. 774 66

We have isolated and characterised the 5' region of a member of the carp myosin heavy chain gene family. Expression of this gene has previously been shown to be induced by an increase in environmental temperature and is restricted to the small-diameter white myotomal muscle fibres which are associated with growth. The whole isoform gene, including potential regulatory sequence 5' to the transcription start site and the 3' untranslated region was cloned in a lambda2001 bacteriophage vector. Studies of the structure of the 5'-end of the gene revealed high amino acid sequence similarity with translated exons 3-7 of mammalian myosin heavy chain genes indicating identical exon/intron boundaries. The overall length of the gene was however only about one half of that in mammals and birds due to shorter introns. The region 5' to the transcription unit was sequenced and revealed the presence of putative TATA and CCAAAT boxes. In order to study the regulation of expression, a series of endonuclease-generated fragments from the 5' flanking sequence were spliced to chloramphenicol acetyltransferase reporter vectors and used in cell transfection assays or direct gene injection into carp skeletal muscle. The 5' flanking region, which contains a consensus sequence known as an E-box (CANNTG) and a MEF2 binding site, was shown to improve the expression of the reporter gene in fish acclimated at 18 degrees C or 28 degrees C. Unlike the coding region, there was little similarity between the 5'-upstream sequence (promoter region) when compared with sequences flanking the 5'-end of the other myosin heavy chain genes in mammals or chicken.
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PMID:The characterisation of the 5' regulatory region of a temperature-induced myosin-heavy-chain gene associated with myotomal muscle growth in the carp. 866 10

Deoxyribonuclease I (DNase I)-like enzyme from the liver of the carp (Cyprinus carpio) was purified to homogeneity and further characterized. Ion exchange chromatography on DEAE-cellulose, molecular filtration on Sephacryl S-300 and Con A-Sepharose affinity chromatography were applied for enzyme isolation. Carp liver DNase, similarly to DNase I from bovine pancreas, was found to be an endonuclease that hydrolyses linear DNA from salmon sperm as well as circular DNA forms--plasmid and cosmid. The purified enzyme is a glycoprotein and shows microheterogeneity, as observed in DNase zymograms prepared after native and two-dimensional electrophoresis (2D-PAGE). The composition of sugar component of the enzyme was characterized. Special attention was focused on the ability of carp liver DNase to interact with carp liver actin. The carp liver enzyme was inhibited by endogenous actin. The estimated binding constant of carp liver DNase to carp liver actin was calculated to be 1.1 x 10(6) M(-1).
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PMID:Carp liver DNase--isolation, further characterization and interaction with endogenous actin. 1562 19