Abstract:
:A novel cytoplasmic gene expression system has been developed. This system differs from other expression systems in that it relies on the co-delivery of plasmid DNA and T7 RNA polymerase (RNAP) during transfection. The plasmid contains a T7 RNAP gene driven by the T7 promoter (T7 autogene) and a functional/reporter gene driven by another T7 promoter (T7T7/T7-gene construct). Once this DNA-enzyme complex is introduced into eukaryotic cells, the transcription of the T7 RNAP and the functional/reporter genes is initiated by the co-delivered T7 RNAP. The T7 RNAP, which is responsible for the initiation and maintenance of expression of both T7 and functional/reporter genes, is replenished by translation of newly synthesized T7 mRNA. This T7 system was designed in such a manner that the expression of the functional/reporter genes can occur in the cytoplasm and does not require any nuclear involvement. When transfected by either a pT7T7/T7Luc or a pT7T7/T7hGH plasmids with the cointroduced T7 RNAP, mouse L cells were found to express high levels of luciferase immediately after transfection, apparently due to the cytoplasmic gene expression; the expression of human growth hormone (hGH) could be sustained for at least 6 days. Both T7 and hGH mRNA were expressed by the cells transfected with pT7T7/T7hGH. These results suggest that this cytoplasmic expression system may be used for certain targets of somatic gene therapy.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Chen X,Li Y,Xiong K,Wagner TEdoi
10.1093/nar/22.11.2114subject
Has Abstractpub_date
1994-06-11 00:00:00pages
2114-20issue
11eissn
0305-1048issn
1362-4962journal_volume
22pub_type
杂志文章abstract::Bacillus stearothermophilus large ribosomal subunits were reconstituted in the presence of 5S rRNAs from different origins and tested for their biological activities. The results obtained have shown that eubacterial and archaebacterial 5S rRNAs can easily substitute for B. stearothermophilus 5S rRNA in the reconstitut...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/16.8.3511
更新日期:1988-04-25 00:00:00
abstract::Accumulation of replicative intermediates of the bacteriophage phi X174 was observed in E. coli C infected cells when phage DNA methylation has been inhibited by nicotinamide or when cells were infected with a temperature-sensitive mutant in gene A. Analysis of the accumulating replicative intermediates by electron mi...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/4.10.3483
更新日期:1977-10-01 00:00:00
abstract::Human transcription factor IIIC contains a 55 kDa polypeptide which specifically interacts with the Adenovirus 2 VAI gene promoter and which mimics most of the DNA binding properties of the entire factor. The specificity and affinity of this protein:DNA interaction was demonstrated by: (i) Separation of purified fract...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/18.16.4743
更新日期:1990-08-25 00:00:00
abstract::The eukaryotic genome evolves under the dual constraint of maintaining coordinated gene transcription and performing effective DNA replication and cell division, the coupling of which brings about inevitable DNA topological tension. DNA supercoiling is resolved and, in some cases, even harnessed by the genome through ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx198
更新日期:2017-06-02 00:00:00
abstract::The nucleotide sequence of the Anacystis nidulans 23S rRNA gene, including the 5'- and 3'-flanking regions has been determined. The gene is 2876 nucleotides long and shows higher primary sequence homology to the 23S rRNAs of plastids (84.5%) than to that of E. coli (79%). The predicted rRNA transcript also shares many...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.7.3373
更新日期:1984-04-11 00:00:00
abstract::We have used in vitro site-directed mutagenesis with synthetic DNA oligonucleotides to introduce single nucleotide mutations in yeast mtDNA. In addition to the expected DNA alterations we also recovered with high frequency mutants with large deletions and insertions which arose through interaction with the synthetic D...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/11.24.8595
更新日期:1983-12-20 00:00:00
abstract::A general procedure, based on a new activated alkyne linker, for the preparation of peptide-oligonucleotide conjugates (POCs) on solid support has been developed. With this linker, conjugation is effective at room temperature (RT) in millimolar concentration and submicromolar amounts. This is made possible since the u...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkr603
更新日期:2011-11-01 00:00:00
abstract::Use of the polymerase chain reaction (PCR) to amplify variable numbers of tandem repeat (VNTR) loci has become widely used in genetic typing. Unfortunately, preferential amplification of small allelic products relative to large allelic products may result in incorrect or ambiguous typing in a heterozygous sample. The ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/23.15.3050
更新日期:1995-08-11 00:00:00
abstract::We describe a database of macromolecular motions meant to be of general use to the structural community. The database, which is accessible on the World Wide Web with an entry point at http://bioinfo.mbb.yale.edu/MolMovDB , attempts to systematize all instances of protein and nucleic acid movement for which there is at...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.18.4280
更新日期:1998-09-15 00:00:00
abstract::Due to the importance of protein phosphorylation in cellular control, many researches are undertaken to predict the kinase-specific phosphorylation sites. Referred to our previous work, KinasePhos 1.0, incorporated profile hidden Markov model (HMM) with flanking residues of the kinase-specific phosphorylation sites. H...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm322
更新日期:2007-07-01 00:00:00
abstract::The molecular mechanism in pancreatic β cells underlying hyperlipidemia and insulin insufficiency remains unclear. Here, we find that the fatty acid-induced decrease in insulin levels occurs due to a decrease in insulin translation. Since regulation at the translational level is generally mediated through RNA-binding ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky867
更新日期:2018-12-14 00:00:00
abstract::Preparation of large amount of single-stranded circular DNA in high selectivity is crucial for further developments of nanotechnology and other DNA sciences. Herein, a simple but practically useful methodology to prepare DNA rings has been presented. One of the essential factors is to use highly diluted T4 ligase buff...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx553
更新日期:2017-09-06 00:00:00
abstract::We report the use of MonoQ FPLC (Fast Protein Liquid Chromatography) for the rapid purification of a novel Type II restriction endonuclease PmaCI, from Pseudomonas maltophila, which recognises the sequence 5'-CAC decreases GTG-3'. The resulting enzyme is free of other nucleases to a level suitable for its characterisa...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.3.1293
更新日期:1986-02-11 00:00:00
abstract::SPRING (http://algorithm.cs.nthu.edu.tw/tools/SPRING/) is a tool for the analysis of genome rearrangement between two chromosomal genomes using reversals and/or block-interchanges. SPRING takes two or more chromosomes as its input and then computes a minimum series of reversals and/or block-interchanges between any tw...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl169
更新日期:2006-07-01 00:00:00
abstract::The effect of UV-irradiation on the conversion of the single-stranded DNA of the parvovirus Minute-Virus-of-Mice (MVM) to duplex Replicative Forms (RF) was studied after infection of mouse A9 fibroblasts. UV-irradiation of the virus prior to infection of unirradiated cells resulted in a dose-dependent, single-hit, inh...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/10.8.2577
更新日期:1982-04-24 00:00:00
abstract::No analogous nucleoside triphosphate was found which acts as well as ATP in binding to and supporting catalysis of leucyl-tRNA synthetase from Escherichia coli MRE 600. However, there are numerous nucleotides which are able to replace ATP, but with lower efficiency. The 6-amino group of the adenine ring and the 2'-hyd...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/3.8.2067
更新日期:1976-08-01 00:00:00
abstract::We examined the 3'-flanking regions required for accurate 3'-end formation of wheat histone H3 mRNA using gene expression in transformed sunflower cells. The introduction of mutations into the conserved sequence AATGGAAATG in the 3'-flanking region of plant histone genes, located 22 bp upstream from the polyadenylatio...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/22.6.1052
更新日期:1994-03-25 00:00:00
abstract::The Gibbs Motif Sampler is a software package for locating common elements in collections of biopolymer sequences. In this paper we describe a new variation of the Gibbs Motif Sampler, the Gibbs Recursive Sampler, which has been developed specifically for locating multiple transcription factor binding sites for multip...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkg608
更新日期:2003-07-01 00:00:00
abstract::A set of programs was developed for searching nucleic acid and protein sequence data bases for sequences similar to a given sequence. The programs, written in FORTRAN 77, were optimized for vector processing on a Hitachi S810-20 supercomputer. A search of a 500-residue protein sequence against the entire PIR data base...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/14.1.57
更新日期:1986-01-10 00:00:00
abstract::Translesion synthesis (TLS) employs low fidelity polymerases to replicate past damaged DNA in a potentially error-prone process. Regulatory mechanisms that prevent TLS-associated mutagenesis are unknown; however, our recent studies suggest that the PCNA-binding protein Spartan plays a role in suppression of damage-ind...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks1267
更新日期:2013-02-01 00:00:00
abstract::Methionyl-tRNA synthetase belongs to the class I aminoacyl-tRNA synthetase family characterized both by a catalytic center built around a Rossmann Fold and by the presence of the two peptidic marker sequences HIGH and KMSKS. In this study, the role of the 21HLGH24 motif of Escherichia coli methionyl-tRNA synthetase wa...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/23.23.4793
更新日期:1995-12-11 00:00:00
abstract::The effect of different levels of mercury substitution on the rate and extent of hybridisation of globin mRNA with a complementary DNA (cDNA) copy has been investigated. It was found that mercuration significantly reduces both the rate of hybridisation and the extent of the reaction, but that these effects are abolish...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/7.8.2357
更新日期:1979-12-20 00:00:00
abstract::Specific targeting of protein–nucleic acid interactions is an area of current interest, for example, in the regulation of gene-expression. Most transcription factor proteins bind in the DNA major groove; however, we are interested in an approach using small molecules to target the minor groove to control expression by...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw1232
更新日期:2017-02-17 00:00:00
abstract::The miRNA Registry provides a service for the assignment of miRNA gene names prior to publication. A comprehensive and searchable database of published miRNA sequences is accessible via a web interface (http://www.sanger.ac.uk/Software/Rfam/mirna/), and all sequence and annotation data are freely available for downloa...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh023
更新日期:2004-01-01 00:00:00
abstract::Nuclei isolated from cultured mouse and rat cell lines initiated pre-rRNA chains at the correct site and with the correct nucleotide specificity (A for mouse, G for rat). Nucleic acid filter hybridization and S1 nuclease mapping were used to analyze the RNA products initiated with nucleoside (beta-S)triphosphates. Ini...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/12.21.8115
更新日期:1984-11-12 00:00:00
abstract::DNA circuit is a versatile and highly-programmable toolbox which can potentially be used for the autonomous sensing of dynamic events, such as biomolecular interactions. However, the experimental implementation of in silico circuit designs has been hindered by the problem of circuit leakage. Here, we systematically an...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkw447
更新日期:2016-08-19 00:00:00
abstract::It has been inferred from DNA sequence analyses that in echinoderm mitochondria not only the usual asparagine codons AAU and AAC, but also the usual lysine codon AAA, are translated as asparagine by a single mitochondrial (mt) tRNAAsn with the anticodon GUU. Nucleotide sequencing of starfish mt tRNAAsn revealed that t...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/27.7.1683
更新日期:1999-04-01 00:00:00
abstract::Establishment and differentiation of mammary alveoli during pregnancy are controlled by prolactin through the transcription factors STAT5A and STAT5B (STAT5), which also regulate temporal activation of mammary signature genes. This study addressed the question whether the methyltransferase and transcriptional co-activ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv776
更新日期:2015-10-15 00:00:00
abstract::We have used 2-aminopurine (2AP) as a fluorescent probe in the template strand of a 13/20mer primer/template (D) to detect deoxynucleoside triphosphates (N)-dependent conformational changes exhibited by RB69 DNA polymerase (ED) complexes. The rates and amplitudes of fluorescence quenching depend hyperbolically on the ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkm587
更新日期:2007-01-01 00:00:00
abstract::Ribosomal protein L11 and its associated binding site on 23S rRNA together comprise one of the principle components that mediate interactions of translation factors with the ribosome. This site is also the target of the antibiotic thiostrepton, which has been proposed to act by preventing important structural transiti...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkh644
更新日期:2004-06-15 00:00:00