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The synthesis of 3 -phosphoadenosine-5 -phosphosulfate (PAPS) from inorganic sulfate and ATP requires two enzymes, ATP sulfurylase (SUL) and adenosine-5 -phosphosulfate kinase (KIN). In bacteria, fungi, yeast and plants, the two enzymes are present on separate polypeptide chains. We have identified the first animal gene coding for these enzymes. In the marine worm, Urechis caupo (Uc), both SUL...
We have developed a system for revealing informative and useful haplotypes for the human Y chromosome using PCR. Variant alphoid satellite DNA subunits were amplified and analysed by digestion with HindIII to score a restriction site polymorphism, or on polyacrylamide gels to reveal 13 heteroduplex haplotypes. Heteroduplexes are double-stranded DNA molecules containing mismatches; the haplotype...
We have isolated four non-tissue-specific helix-loop-helix (HLH) transcription factors from Xenopus laevis (Xl). While some are clearly orthologous to known mammalian HLH proteins, others have dramatic amino-acid differences in otherwise highly conserved protein domains. We propose that these changes arose following the tetraploidization of Xl approx. 30 Myr ago.
A cDNA encoding one of two major allotypic forms of bovine pancreatic preprocarboxypeptidase A (preproCPA) has been cloned, and its entire nucleotide (nt) sequence determined. The cloned cDNA contains a 26-nt 5 -noncoding region, a 1260-nt open reading frame and a 51-nt 3 -noncoding region. The amino acid (aa) sequence deduced from the gene sequence contains Ile 179 , Ala ...
A detailed restriction map of the murine TGF-β1 locus (encoding transforming growth factor β1) was established and the precise junctional sequences of its seven exons and corresponding introns were elucidated. While the exons ranged in size from 78 to 357 bp, the introns ranged from about 1 to 6.5 kb. The promoter-proximal segment contains many putative regulatory motifs which may dictate TGF-β1...
We previously reported the identification of the human CAP and CAP2 genes which encode proteins related to the yeast adenylyl cyclase (CYR)-associated CAP protein. The rat CAP homolog, MCH1, has also been previously cloned. We have cloned a cDNA encoding the rat homolog of CAP2. Rat CAP/MCH1 and CAP2 are 63% identical to each other. Using the reverse transcription-polymerase chain reaction (RT-PCR)...
Two Saccharomyces cerevisiae proteins of 21 and 27 kDa co-purify with a novel enhancer of Ga14p DNA binding activity (Egdp) [Parthun et al., Mol. Cell. Biol. 12 (1992) 5683-5689]. Mutations in the EGD1 gene encoding the 21-kDa protein (Egd1p) have been shown to affect the kinetics and extent of the Ga14p-mediated, galactose-induced activation of the GAL genes. Egd1p is homologous to human BTF3b,...
The 5 cap (m 7 GpppN) and the poly(A) tail of eukaryotic mRNAs work in concert to establish an efficient level of translation in vivo. Nevertheless, several mRNAs naturally lack a cap or a poly(A) tail. Determining how these messages effectively compete for the translational machinery not only reveals alternative mechanisms for translational competence, but can also underscore similarities...
Two cDNAs encoding calnexin (Cln)-like and calreticulin (Crl)-like proteins have been isolated by immunoscreening of a maize leaf cDNA library. In the deduced amino acid (aa) sequences, several regions that are conserved for Cln and Crl proteins from all sources have been identified. These regions can be classified into two distinct motifs which are repeated four times each in Cln and three times...
Polymerase chain reaction amplification of the cDNA encoding steroid 17α-hydroxylase (P450c17) demonstrated very low levels of this transcript and a shorter variant of still lower abundance in rat adrenals and brain. Sequence analysis of the two amplified products revealed that the shorter variant resulted from a deletion of the second exon which does not maintain the open reading frame, suggesting...
Methenyltetrahydrofolate synthetase (MTHFS) catalyses the obligatory initial metabolic step in the intracellular conversion of 5-formyltetrahydrofolate to other reduced folates. We have isolated and sequenced a human MTHFS cDNA which is 872-bp long and codes for a 203-amino-acid protein of 23 229 Da. Escherichia coli BL21 (DE3), transfected with pET11c plasmids containing an open reading frame encoding...
We identified and cloned the rabbit SP-B gene, encoding the pulmonary surfactant-associated protein, and sequenced its upstream region from -2635 to +428, including a much larger fragment of the upstream region than has previously been reported for an SP-B for any species. Rabbit SP-B showed substantial homology to its human counterpart in the coding and noncoding regions immediately upstream from...
We have isolated the rat bone marrow stromal cell antigen 1-encoding cDNA (BST-1) from a pancreatic islet cDNA library. The cDNA encodes a 319-amino-acid (aa) protein whose aa sequence shows homology with mammalian CD38 (33%), Aplysia ADP-ribosyl cyclases (33%), as well as mouse (86%) and human (72%) BST-1.
A linear 5.2-kb HS2/β-globin construct with an upstream KpnI terminus (4-nucleotide (nt) 3 protruding single strand, PSS) and a downstream SalI terminus (4-nt 5 PSS) was microinjected into fertilized mouse eggs. The injected DNA fragments integrated into the mouse genome primarily as a head-to-tail tandem array. Chromosome/transgene junctions were obtained from seven of eight transgenic animals...
A novel cDNA clone, Oshsp18.0 cDNA, encoding a rice (Oryza sativa L. cv. Tainong 67) 18.0-kDa heat-shock protein (HSP), was isolated from a cDNA library of heat-shocked rice seedlings by use of the rice HSP cDNA, Oshsp7.3 cDNA, as a probe. The sequence showed that Oshsp18.0 cDNA contains a 749-bp insert encoding an ORF of 160 amino acids, with a predicted molecular mass of 18.0 kDa and a pI of 7...
The Candida glabrata (Cg) TRP1 and HIS3 genes have been isolated by complementation of the Saccharomyces cerevisiae (Sc) trp1 and his3 mutants, respectively. Cg TRP1 encodes a polypeptide of 217 amino acids (aa), whose aa sequence is 58% identical to that of Sc TRP1. Cg HIS3 encodes a polypeptide of 210 aa, whose aa sequence is 73% identical to that of the Sc HIS3. Both Cg TRP1 and HIS3 were disrupted...
The flounder (Paralichthys olivaceus) growth hormone (GH)-encoding gene (fGH) and its promoter region were cloned and sequenced following amplification of genomic DNA by the polymerase chain reaction. The fGH gene is 2.1-kb long and consists of six exons and five introns. In the 5 -flanking region of the determined transcription start point, a potential TATA box is located at -24, and Pit-1/GHF-1-binding...
A novel method is described for the rapid concentration of particular cDNAs and their mapping to specific regions of a genome. The strategy for cDNA scanning is based on the hybridization of an entire library of cDNAs to a large fragment of genomic DNA that is covalently bound to latex particles. The hybridized cDNAs are eluted, amplified by PCR and cloned into a λ vector. Selected cDNAs that...
A 5 truncated caprine (ca) -casein-encoding gene ( Cas) was fused to the 3 end of a 3 truncated ca βCas. The Cas form comprised the 0.8-kb 3 end of intron 2, the remaining part of the transcription unit containing codon -- 2 to stop 172, and 0.43 kb of the 3 flanking region. The βCas form comprised a 3-kb 5 flanking region and the 5 end of the transcription unit terminating 69...
We have identified a gene encoding the human ribosomal protein (r-protein) S7. The S7 gene contains seven exons and six introns spanning about 6 kb. Organization of the gene is similar to that of Xenopus laevis S8, the only homologous intron-containing gene isolated so far. An mRNA transcribed from this gene has an open reading frame (ORF) of 582 necleotides (nt), which encodes a protein of 194...
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