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The cDNA coding for GT3-synthase has been cloned from a rat fetal brain cDNA library. The complete nucleotide sequence and the deduced amino acid sequence of the rat GT3-synthase cDNA were highly homologous to those of the mouse GT3-synthase. Quantitative RT-PCR showed that, as a key enzyme for the synthesis of c -series gangliosides, the expression of GT3-synthase was developmentally regulated...
Overexpression of αB-crystallin is associated with numerous neurodegenerative diseases and abnormal cell growth patterns. To study the mechanisms involved in the control of the transcriptional activity of the gene we have characterized its expression in different chicken tissues. The sequence of the αB-crystallin cDNA isolated from chicken testis and 6-day-old chick embryo is highly homologous to...
As a step toward understanding the transcriptional regulation of the adrenocorticotropin receptor (ACTH-R) gene, we examined the full length cDNA sequence of the mouse ACTH-R by rapid amplification of cDNA ends, and the organization of the gene. Mouse ACTH-R mRNA consists of 374 bp in the 5 -untranslated region (UTR), 888 bp in the coding sequence, and 445 bp in the 3 -UTR, the 1707 bp being fairly...
A putative promoter element responsible for transcription of the spliced leader (SL) gene of Trypanosoma cruzi was identified by overlapping deletion and linker scanning analyses of the upstream flanking sequences using the bacterial chloramphenicol acetyltransferase (CAT) gene as a reporter in transient transfections of cultured epimastigotes. Deletion or substitution of a proximal sequence element...
Inosine-containing degenerate PCR primers corresponding to the heme-binding domain of cytochrome P450 proteins have been synthesized and used for cloning cDNAs by the RT-PCR technique from Solanum chacoense. One clone in which the primer was immediately followed by sequences corresponding to the remaining part of the conserved domain was obtained. A leaf cDNA and a genomic library were constructed...
We have previously revealed that in the brine shrimp Artemia franciscana an AluI DNA family of repeats, 113 bp in length, is the major component of the constitutive heterochromatin and that this repetitive DNA shows a stable curvature that confers a solenoidal geometry on the double helix in vitro. It was suggested that this particular structure may play a relevant role in determining the condensation...
Previous work on the promoter region of the 63 kDa keratin gene demonstrated that in vivo footprints did not change during the transition from low-level to high-level transcription. Reverse transcription polymerase chain reaction and in vivo footprinting were used to determine whether these DNA-protein interactions are present before transcription begins. The results presented indicate that during...
The bacterial cloning vector, pGreenscript A, derived from the mutated Aequorea green fluorescent protein (GFP-S65A) gene, when expressed in E. coli produced colonies that showed yellow color under daylight and strong green fluorescence under long-wave ultraviolet light. The vector was used to select for inserted foreign genes based on the loss of the yellow color/green fluorescence of E. coli cells...
chromokinesin is a developmentally down-regulated gene with specific expression in proliferating cells during embryonic chick development. It encodes a DNA-binding motor protein localized along the chromosome arm during mitosis, suggesting that the protein may be a component of the long-observed, yet poorly understood `ejection force' hypothesized to be involved in controlling the direction and speed...
Two novel shuttle vectors for mycobacteria are described which have been derived from the expression system pSD5 developed in our laboratory. Plasmid pSD5B is a promoter-selection vector containing a promoterless lacZ gene and allows the identification of mycobacterial promoters by the blue colour of the colonies on solid media containing XGal. Moreover, the chronological order of appearance of blue...
We isolated genomic clones for bovine prostaglandin (PG) F 2α receptor by the standard plaque hybridization method, using the cDNA fragments of bovine PGF 2α receptor (PGF 2α R) as probe DNAs. The coding regions of this receptor gene were interspersed by a large intron sequence (33 kb) at the splice junction in the sixth transmembrane domain. The 5 -RACE experiments...
We report the cloning and characterisation of a cDNA that encodes a novel member of the Xenopus laevis 14-3-3 protein family. Sequence analysis reveals that the cDNA-encoded protein shares 84% identity with the rat, human or sheep 14-3-3ζ isoform, and between 66% and 77% identity with bovine, human or rat β, bovine γ, human τ, Drosophila 14-3-3 and a previously isolated Xenopus member. The corresponding...
We have purified the chromosomal high mobility group (HMG) protein HMGd from maize suspension culture cells, determined the N-terminal amino acid (aa) sequence, and isolated the corresponding cDNA. Sequence analysis showed that the cDNA encoded a protein of 126 aa residues with a theoretical mass of 14 104 Da. The protein contains an HMG-box DNA-binding domain and a short acidic C-terminal tail...
Large samples of original cDNAs encoding neonatal and adult mouse fast skeletal muscle troponin T (fTnT) have been isolated and characterized. The results demonstrate expression relationships of 8 alternatively spliced exons of the fTnT gene and reveal the primary structure of as many as 13 fTnT isoforms that diverge into acidic and basic classes due to differential mRNA splicing in the N-terminal...
To molecularly characterize the influence of external pH in the secretion of enzymes by filamentous fungi, we have cloned and sequenced the palF gene of Aspergillus nidulans (An). An An wild-type (wt) chromosome VII-specific cosmid library was used to transform a palF15 mutant strain. Selection for complementation was done on medium containing β-glycerolphosphate as the sole P i source. Two...
The two full-length cDNAs encoding ESG1 (Enhancer of split groucho) and related AES (Amino Enhancer of split) proteins of 767 and 197 amino acids, respectively, were cloned and sequenced from the African frog Xenopus laevis. The amino acid sequence of Xenopus ESG1 protein had 61% identity to the full-length Drosophila groucho. Xenopus AES protein exhibited 91%, 58% and 48% identity to the mouse AES,...
A 2.5kb homeobox (HB)-containing cDNA (hahr1) was isolated from a library prepared from rootlets of Helianthus annuus using a polymerase chain reaction (PCR)-based strategy. The putative protein product (77kDa) contains the homeodomain (HD) and an acidic domain at the N-terminal region (residues 72-155). The deduced amino acid sequence of hahr1 shares a 53% sequence identity with GLABRA2, a HD protein...
Smooth muscle cells (SMC) express a battery of cell-restricted differentiation genes, many of which are down-regulated during the course of vascular disease. Here, we present the mRNA expression, genomic structure and chromosomal mapping of the gene encoding human smooth muscle cell calponin (SMCC). Human SMCC transcripts are restricted to tissues and cells of SMC origin and, in the latter case, appear...
Glycogen storage disease type III (GSD-III) is caused by a deficiency of glycogen debranching enzyme (AGL) activity. Patients are found to have deficient AGL activity in both muscle and liver, and also enzyme deficiency in the liver, but not in muscle. To determine the molecular basis of enzymatic variability in GSD-III and to elucidate the mechanism for control of tissue-specific expression of AGL,...
The myeloperoxidase (MPO) gene is expressed specifically in immature myeloid cells. The MPO gene includes a promoter proximal enhancer which is coincident with DNaseI hypersensitive chromatin sites and is specifically active in myeloid cell lines. We developed transgenic murine lines in which 1.3kb of murine MPO proximal 5 flanking region DNA was linked to a TATAA homology and RNA initiation site...
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