High frequency recombination betweenloxP sites in human chromosomes mediated by an adenovirus vector expressing Cre recombinase

1995 ◽  
Vol 21 (6) ◽  
pp. 429-441 ◽  
Author(s):  
Ping Wang ◽  
Martina Anton ◽  
Frank L. Graham ◽  
Silvia Bacchetti
Keyword(s):  
High Frequency ◽  
Adenovirus Vector ◽  
Cre Recombinase ◽  
Human Chromosomes

scholarly journals High-Efficiency System for the Construction of Adenovirus Vectors and Its Application to the Generation of Representative Adenovirus-Based cDNA Expression Libraries

2006 ◽  
Vol 80 (11) ◽  
pp. 5435-5450 ◽  
Author(s):  
Moritz Hillgenberg ◽  
Christian Hofmann ◽  
Herbert Stadler ◽  
Peter Löser
Keyword(s):  
Human Liver ◽  
Recombinant Adenovirus ◽  
Adenovirus Vector ◽  
Cre Recombinase ◽  
Cdna Expression Library ◽  
Packaging Signal ◽  
Expression Library ◽  
Cdna Expression ◽  
Recombinant Adenovirus Vector ◽  
Adenovirus Vectors

ABSTRACT We here describe a convenient system for the production of recombinant adenovirus vectors and its use for the construction of a representative adenovirus-based cDNA expression library. The system is based on direct site-specific insertion of transgene cassettes into a replicating donor virus. The transgene is inserted into a donor plasmid containing the viral 5′ inverted terminal repeat, the complete viral packaging signal, and a single loxP site. The plasmid is then transfected into a Cre recombinase-expressing packaging cell line that has been infected with a donor virus containing a partially deleted packaging signal flanked by loxP sites. Cre recombinase, by two steps of action, sequentially catalyzes the generation of a nonpackageable donor virus acceptor substrate and the generation of the desired recombinant adenovirus vector. Due to its growth impairment, residual donor virus can efficiently be counterselected during amplification of the recombinant adenovirus vector. By using this adenovirus construction system, a plasmid-based human liver cDNA library was converted by a single step into an adenovirus-based cDNA expression library with about 106 independent adenovirus clones. The high-titer purified library was shown to contain about 44% of full-length cDNAs with an average insert size of 1.3 kb. cDNAs of a gene expressed at a high level (human α1-antitrypsin) and a gene expressed at a relatively low level (human coagulation factor IX) in human liver were isolated from the adenovirus-based library using an enzyme-linked immunosorbent assay-based screening procedure.

Efficient Sequential Gene Regulation via FLP- and Cre-Recombinase Using Adenovirus Vector in Mammalian Cells Including Mouse ES Cells

2006 ◽  
Vol 50 (10) ◽  
pp. 831-843 ◽  
Author(s):  
Saki Kondo ◽  
Yuzuka Takahashi ◽  
Seiji Shiozawa ◽  
Hirotake Ichise ◽  
Nobuaki Yoshida ◽  
...  
Keyword(s):  
Gene Regulation ◽  
Mammalian Cells ◽  
Es Cells ◽  
Adenovirus Vector ◽  
Cre Recombinase ◽  
Mouse Es Cells

Digestion of human chromosomes by means of the isoschizomers MspI and HpaII

Genome ◽  
1994 ◽  
Vol 37 (5) ◽  
pp. 770-774 ◽  
Author(s):  
A. M. Fernandez-Peralta ◽  
P. Navarro ◽  
I. Tagarro ◽  
J. J. Gonzalez-Aguilera
Keyword(s):  
Mechanism Of Action ◽  
Satellite Dna ◽  
High Frequency ◽  
Chromosome Region ◽  
Cpg Methylation ◽  
Restriction Endonucleases ◽  
High Rate ◽  
Human Chromosomes ◽  
Banding Patterns ◽  
Insight Into

The isoschizomers MspI and HpaII are four base cutter (C↓CGG) restriction endonucleases, HpaII being sensitive to methylation of the internal cytosine. Human chromosomes treated with MspI have produced inconsistent results between laboratories, while HpaII has always been described as a nonbanding enzyme when used on human chromosomes. These results have been explained on the basis of both rarity of the CpG doublet in vertebrate genomes and high rate of CpG methylation (5mCpG). We demonstrated consistent banding patterns subsequent to digestions with MspI and HpaII. On euchromatin, MspI (but not HpaII) digests the DNA of R regions and thus R-bands apparently contain many more CCGG sites (mostly methylated) than G-bands. In heterochromatin, extensive digestion of the 9q12 region not only by MspI but also by HpaII reveals a heterochromatic domain with a high frequency of unmethylated CCGG sites, most probably within the satellite 3 DNA fraction. In addition, enzymatic digestions of the Yq12 heterochromatin, when this region is undercondensed by 5-azacytidine, contribute to deepen the insight into the mechanism of action of this cytidine analog and at the same time reinforce the idea of the heterogeneity of this chromosome region where domains with unmethylated CCGG sites may also exist.Key words: human chromosomes, methylation, restriction endonucleases, heterochromatin, satellite DNA.

The Microstructure of Steatite (Protoenstatite)

1985 ◽  
Vol 43 ◽  
pp. 236-237
Author(s):  
W. E. Lee ◽  
A. H. Heuer
Keyword(s):  
High Frequency ◽  
Glass Ceramics ◽  
Magnesium Silicate ◽  
Beam Current ◽  
Glassy Matrix ◽  
Angle Of Incidence ◽  
X Ray ◽  
Mechanical And Electrical Properties ◽  
Argon Ions ◽  
High Temperature Polymorph

IntroductionTraditional steatite ceramics, made by firing (vitrifying) hydrous magnesium silicate, have long been used as insulators for high frequency applications due to their excellent mechanical and electrical properties. Early x-ray and optical analysis of steatites showed that they were composed largely of protoenstatite (MgSiO3) in a glassy matrix. Recent studies of enstatite-containing glass ceramics have revived interest in the polymorphism of enstatite. Three polymorphs exist, two with orthorhombic and one with monoclinic symmetry (ortho, proto and clino enstatite, respectively). Steatite ceramics are of particular interest a they contain the normally unstable high-temperature polymorph, protoenstatite.Experimental3mm diameter discs cut from steatite rods (∼10” long and 0.5” dia.) were ground, polished, dimpled, and ion-thinned to electron transparency using 6KV Argon ions at a beam current of 1 x 10-3 A and a 12° angle of incidence. The discs were coated with carbon prior to TEM examination to minimize charging effects.

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