Search results for "Cloning"
showing 10 items of 498 documents
Differential Regulation of CCL22 Gene Expression in Murine Dendritic Cells and B Cells
2005
Abstract The activated T cell-attracting CC chemokine CCL22 is expressed by stimulated B cells and mature dendritic cells (DC). We have cloned and sequenced the complete mouse gene, including 4 kb of the 5′-flanking promoter region, and detected two distinct sites for initiation of transcription by 5′-RACE. Reporter gene assays indicate that the promoter reflects the specificity of the endogenous gene. Within the proximal promoter region, we identified potential binding sites for NF-κB, Ikaros, and a putative GC box. All three regions bind proteins. The NF-κB site was shown to specifically bind NF-κB subunits p50 and p65 from nuclear extracts of LPS-stimulated B cells, B cell line A20/2J, T…
Chromatin structure of transposon Tn903 cloned into a yeast plasmid
1989
Transposon Tn903 contains the APH gene for kanamycin resistance, which is active in yeast [A. Jiménez and J. Davies (1980) Nature (London) 287, 869-871] and is flanked by two inverted repeats (IR) 1057 bp long. When plasmid pAJ50, carrying Tn903 and the 2-microns circle origin of replication, is cloned into Saccharomyces cerevisiae, nucleosomes are assembled in vivo on the prokaryotic DNA of the transposon. Indirect end labeling revealed that three nucleosomes are preferentially positioned on symmetrical sequences from both IRs. DNase I digestion also confirmed that the chromatin structure is symmetrical in both IRs. This suggests that sequence determinants are decisive for chromatin struct…
Insertion mutation inactivates the expression of the recessive oncogene lethal(2)giant larvae of Drosophila melanogaster
1986
The Drosophila recessive oncogene lethal(2)giant larvae is located at the extreme left end of the second chromosome close to telomeric repetitive sequences. Of the 20 l(2)gl mutant alleles isolated from wild flies in widespread populations of the Soviet Union and California, all but two appear to represent large deletions which have removed the telomeric repetitive sequences and l(2)gl single copy sequences (Mechler et al. 1985). We have analyzed the structure of the two exceptions: the l(2)glGB52 mutation results from the insertion of a single transposable element of the B104 or roo family, whereas the more complex rearrangements of the l(2)glDV275 mutation consists of an 8 kb interstitial…
The sex determining region of Chironomus thummi is associated with highly repetitive DNA and transposable elements.
1993
The dominant male sex determiner in chromosome III of the midge Chironomus thummi thummi is closely linked to a large cluster of tandem-repetitive DNA elements, the Cla elements, which are otherwise highly repetitive and distributed over more than 200 sites on all chromosomes. Chromosome III displays a hemizygous cluster of Cla elements in males but not in females. The chromosomal location of this hemizygous Cla element cluster is in the region of the male determiner M as localized by cytogenetic analysis. With Cla elements as hybridization probe, it was possible to clone a large part of the sex determining region. Molecular analysis of the DNA of males and females in this region displayed …
P sequences ofDrosophilla Subobscuralack exon 3 and may encode a 66 kd repressor-like protein
1991
Abstract Several P homologous sequences have been cloned and sequenced from Drosophila subobscura. These sequences are located at the 85DE region of the O chromosome and at least three of them are organized in tandem. We have identified four copies which exhibit strong similarity between them. All of the isolated elements are truncated at the 5' and 3' ends. They have lost the inverted terminal repeats and exon 3, but maintain exons 0, 1 and 2. They are transcribed producing a polyadenylated RNA. The structure of these transcripts suggests that they are able to encode a 66 kd repressor-like protein, but not a functional transposase. We ask about the biological role of a potential repressor …
Common genomic structure for the Lepidoptera cadherin-like genes.
2005
A cadherin-like protein present in the midgut epithelial cells of Lepidoptera is associated with insect resistance to Bacillus thuringiensis Cry toxins. We describe for the first time the genes that encode the cadherin-like proteins in Ostrinia nubilalis, Helicoverpa armigera, and Bombyx mori, and analyze their organization. These genes encompass 19.6 kb, 20.0 kb, and 41.8 kb of genomic DNA, respectively, and despite the size heterogeneity, they are all composed of 35 exons that are linked by 34 introns. In contrast to the high variability noted for the sizes of the introns, the sizes of the coding exons were almost completely preserved among the three species, because the intronic sequence…
The Mu1 transposable element of maize contains two promoter signals recognized by the Escherichia coli RNA polymerase.
1990
The galactokinase (GalK) expression plasmid vector system pKO-1 has been used to screen for promoter elements in the maize transposable element Mu1 that function in Escherichia coli. Two transcriptional start points, named S1 and S2, were identified, which are located in the two direct repeats of the transposable element. This paper demonstrates that sequence elements exist in a plant transposable element which function as prokaryotic promotors.
bilbo, a non-LTR retrotransposon of Drosophila subobscura: a clue to the evolution of LINE-like elements in Drosophila
1997
We used the repetitive character of transposable elements to isolate a non-LTR retrotransposon in Drosophila subobscura. bilbo, as we have called it, has homology to TRIM and LOA elements. Sequence analysis showed a 5' untranslated region (UTR), an open reading frame (ORF) with no RNA-binding domains, a downstream ORF that had structural homology to that of the I factor, and, finally, a 3' UTR which ended in several 5-nt repeats. The results of our phylogenetic and structural analyses shed light on the evolution of Drosophila non-LTR retrotransposons and support the hypothesis that an ancestor of these elements was structurally complex.
Quasispecies dynamics and molecular evolution of human norovirus capsid P region during chronic infection.
2009
In this novel study, we have for the first time identified evolutionarily conserved capsid residues in an individual chronically infected with norovirus (GGII.3). From 2000 to 2003, a total of 147 P1-1 and P2 capsid sequences were sequenced and investigated for evolutionarily conserved and functionally important residues by the evolutionary trace (ET) algorithm. The ET algorithm revealed more absolutely conserved residues (ACR) in the P1-1 domain (47/53, 88 %) as compared with the P2 domain (86/133, 64 %). The capsid P1-1 and P2 domains evolved in time-dependent manner, with a distinct break point observed between autumn/winter of year 2000 (isolates P1, P3 and P5) and spring to autumn of y…
Construction and expression of a dual vector for chemo-enzymatic synthesis of plant indole alkaloids inEscherichia coli
2010
A dual vector (pQE-70-STR1-SG) containing coding regions of strictosidine synthase (STR1, EC 4.3.3.2) and strictosidine glucosidase (SG, EC 3.2.1.105) from the Indian medicinal plant Rauvolfia serpentina was constructed. Functional expression of the vector in Escherichia coli cells (M15 strain) was proven by isolation of prepurified enzyme extracts, which show both STR1 and SG activities. Incubation of the enzyme in the presence of tryptamine and secologanin delivered the indole alkaloid cathenamine, demonstrating functional co-expression of both STR1- and SG-cDNAs. Cathenamine reduction by sodium borohydride leading to tetrahydroalstonine revealed the chemo-enzymatic indole alkaloid synthe…