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Query: EC:3.1.30.1 (
S1 nuclease
)
3,660
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
We cloned an ornithine decarboxylase antizyme-encoding gene (Oaz) from a rat liver genomic library. The entire gene was located on a 4367-bp EcoRI fragment, which corresponded to one of two fragments hybridizable with the antizyme-encoding cDNA, Z1, on Southern blot analysis. Sequence analysis of the cloned gene showed that it consisted of five exons which were identical with the cDNA. The transcription start points of the Oaz mRNA were located 75 and 76 nucleotides upstream from the first ATG codon, as determined by
S1 nuclease
protection and primer extension analyses. The 5'-flanking region of the gene contained typical promoter motifs, such as a TATA box and Sp1-binding sites. Introduction of a chimeric gene consisting of the 5'-flanking region and the bacterial cat gene into Chinese hamster ovary cells revealed a promoter activity in the region, which was comparable in strength to that of the simian virus 40 promoter. In addition, we isolated a 12-kb EcoRI fragment, the other sequence hybridizable to the cDNA. Sequence analysis showed that it represented a processed Oaz
pseudogene
and was not able to encode any active protein.
...
PMID:Cloning and characterization of a rat gene encoding ornithine decarboxylase antizyme. 157 40
An EMBL4 recombinant phage which encodes one of the full length of the aphid ribosomal DNA has been isolated from the aphid genomic library. Determination of the complete nucleotide sequence of the aphid 18S rRNA gene revealed that it is 2469 bp with a G + C content of 59%. The aphid 18S rRNA gene studied here is the longest and has the highest G + C content among the 18S rRNA genes examined so far. Evidence provided by the
S1 nuclease
assay suggests that the aphid 18S rRNA gene examined in this study is not a
pseudogene
containing an insertion sequence. Based on the nucleotide sequence of the 18S rRNA gene, we constructed a presumed secondary-structure model of the aphid 18S rRNA. In the aphid 18S rRNA, the eucaryote-specific E21 and 41 region are supposed to be longer and more complex than the counterparts of other 18S rRNA.
...
PMID:The longest 18S ribosomal RNA ever known. Nucleotide sequence and presumed secondary structure of the 18S rRNA of the pea aphid, Acyrthosiphon pisum. 176 96
A total of 29 human genomic DNA clones that hybridize with cDNAs for the sheep and rat Na,K-ATPase beta subunits have been isolated, classified by restriction endonuclease mapping and Southern blot hybridization analysis, and sequenced. One class of clones, designated ATP1BL1, represents a processed
pseudogene
for the beta subunit. The second class, designated ATP1B, includes 15 overlapping genomic clones and represents a functional gene for the human Na,K-ATPase beta subunit. ATP1B spans about 26.7 kb of genomic DNA and includes 24 kb of intron sequence. The complete mRNA transcript for the human beta subunit is encoded by six exons, ranging in size from 81 to 1427 bp. Primer extension and
S1 nuclease
protection experiments with human kidney RNA indicate the presence of two major transcription initiation sites at -510 and -201 to -191, with minor initiation sites at -268, -182 to -174, and -142. The distal initiation site at -510 is preceded by consensus sequences for CAAT and TATA boxes. The DNA sequence preceding the proximal heterogeneous initiation sites contains a CAAT box, but no TATA box. Two of the 12 GC boxes (GGCGGG and CCCGCC) located in the 5' region of ATP1B are located between this CAAT box and the proximal clusters of transcription initiation sites.
...
PMID:Characterization of two genes for the human Na,K-ATPase beta subunit. 255 24
A genomic library of Brugia malayi was constructed and screened by hybridization with a cDNA clone corresponding to a potentially protective antigen of 63 kDa. The antigen is encoded by a multicopy gene family. Five distinct gene copies were isolated and one was characterized in detail by nucleotide sequence analysis. An apparent
pseudogene
was also characterized. The organization of genes encoding the antigen is typical of higher eukaryotes in exon/intron organization although the introns have an unusually high A+T content (75%). Organization of the genomic sequence along with
S1 nuclease
and primer extension analyses indicate that a short untranslated exon is spliced to the 5' end of the mRNAs encoding the antigen.
...
PMID:A multi-copy gene encodes a potentially protective antigen in Brugia malayi. 284 May 77
The POMC gene is predominantly expressed in the pituitary gland; it is also expressed in various extrapituitary tissues. While POMC mRNAs of similar size (approximately equal to 1000 nucleotides) are present in the anterior and neurointermediate lobes of the pituitary, other POMC-expressing tissues contain POMC mRNAs of different sizes. Longer POMC mRNAs are observed in the hypothalamus. Using
S1 nuclease
mapping and mRNA deadenylation by RNase H, we have shown that these large hypothalamic POMC mRNAs have longer poly(A) tails than pituitary POMC transcripts but contain the same transcripted sequences. In contrast, the testes contain POMC transcripts which are smaller than pituitary POMC mRNA. RNase and
S1 nuclease
mapping analyses suggest that these short transcripts do not contain sequences transcribed from pituitary exons 1 and 2. Indeed, as revealed by primer-extension experiments, these transcripts appear to initiate within exon 3 sequences of the POMC gene. The heterogeneous 5'-ends of these short testicular transcripts map into the NH2-terminal portion of the precursor in the region encoding gamma MSH; if ever translated, these transcripts would produce a form of POMC that would be truncated at the NH2-terminus and therefore would be devoid of any signal peptide sequence. Interestingly, the sequence of the short testicular transcripts corresponds to that of the mouse POMC
pseudogene
, suggesting that this POMC
pseudogene
may have derived from genomic integration of testicular transcripts via a cDNA intermediate.
...
PMID:Unusual proopiomelanocortin ribonucleic acids in extrapituitary tissues: intronless transcripts in testes and long poly(A) tails in hypothalamus. 285 1
The primary structure of P31, a 13.300 kDa mammalian 'housekeeping' protein, was deduced from the nucleotide sequence of a rat recombinant cDNA clone. The P31 mRNA has 950 nucleotides and codes for a protein of 121 amino acids. This mRNA is present in several mouse tissues and in other mammals. Nuclei of rapidly growing cells contain mature P31 mRNA and a distinctive set of larger transcripts. P31 is encoded by a multigene family. Five members of the family were isolated from a mouse genomic library and restriction mapping and
S1 nuclease
analysis of four of them (P31-4, P31-12, P31-13 and P31-14) suggest that they are processed genes. The very low homology of the fifth (P31-8) with respect to the corresponding cDNA sequence indicates that it is a
pseudogene
. Although the features of the mRNA and the genes encoding P31 exhibit extensive similarity with those of ribosomal proteins, neither the identity nor the biological function of the protein has yet been determined.
...
PMID:P31, a mammalian housekeeping protein encoded by a multigene family containing a high proportion of pseudogenes. 299 4
A rabbit liver P-450-like
pseudogene
has been isolated from a lambda phage genomic library. Sequence analysis revealed structural homology with respect to the rat P-450b and P-450e genes as well as a similar intron-exon organization. A 5'-proximal TATA box-like sequence and two 3'-distal putative polyadenylation signals were identified, and all putative intron-exon boundaries except at the 3'-splice site of intron 2 were found to follow the GT/AG rule. With allowance for apparent deletions and insertions, the structural homology of the amino acid sequence deduced from the
pseudogene
with respect to rabbit P-450 isozyme 2 is lower for exons 1 through 4 (18-28%) than for exons 5 through 9 (42-65%).
S1 nuclease
mapping showed that mRNAs complementary to the DNA sequence of exon 9 are expressed. However, due to the alterations in the
pseudogene
, it appears that functional P-450 would not be produced from such mRNAs.
...
PMID:Isolation and characterization of a novel cytochrome P-450-like pseudogene. 300 50
We report here a new type of peculiar repetitive sequence, A15T(TC)9T12, which was detected at 750 base pairs (bp) upstream of a rat calmodulin processed
pseudogene
by DNA sequencing of cloned DNA fragments. This sequence element could possibly form a cruciform structure with a 12-AT-pair stem, exposing (CT)9 sequences as a loop.
S1 nuclease
protection experiments failed to identify this element as a cruciform structure but instead detected an alternating purine pyrimidine tract at 50 bp downstream of this element. Total genomic Southern blotting showed that the rat genome contains only a few of these elements.
...
PMID:A peculiar repetitive sequence in the rat genome. 302 31
As shown by Southern blot analysis, the metallothionein-1 (MT-1) genes in rats comprise a multigene family. We present the sequence of the MT-1 structural gene and compare its features with other metallothionein genes. Three MT-1 pseudogenes which we sequenced apparently arose by reverse transcription of processed mRNA transcripts. Two of these, MT-1 psi a and MT-1 psi c, are retrogenes which derive from the MT-1 mRNA, having diverged from the MT-1 gene 6.9 and 2.6 million years ago, respectively. The third, MT-1 psi b, differs from the MT-1 cDNA by only three nucleotide alterations. Surprisingly, MT-1 psi b also preserves sequence homology for 142 base pairs 5' to the transcription initiation site of the parent gene; it contains a promoter sequence sufficient for specifying metal ion induction. We identified, by
S1 nuclease
mapping, an RNA polymerase II initiation site 432 base pairs 5' of the MT-1 transcription initiation site of the MT-1 structural gene which could explain the formation of the mRNA precursor to this
pseudogene
. We were unable to detect MT-1 psi b transcripts, either in liver tissue or after transfection. We conclude that the absence of detectable transcripts from this
pseudogene
is due to either a reduced level of transcription or the formation of unstable transcripts as a consequence of the lack of a consensus sequence normally found 3' of transcription termination in the MT-1 structural gene.
...
PMID:Rat metallothionein-1 structural gene and three pseudogenes, one of which contains 5'-regulatory sequences. 302 30
A processed
pseudogene
for human ceruloplasmin has been isolated that contains DNA corresponding to the functional gene sequence encoding the carboxy-terminal 563 amino acid residues and the 3' untranslated region. The
pseudogene
appears to have arisen from a processed RNA species, since intervening sequences coincident with those of the functional gene have been removed, with the exception of a short segment of intronic sequence which denotes the 5' boundary of the
pseudogene
. The nucleotide sequence of the
pseudogene
is highly homologous (97% sequence identity) with that of the wild-type gene, suggesting that
pseudogene
formation was a relatively recent evolutionary event. In addition to single base substitutions, there is a large 213 base pair (bp) deletion in the
pseudogene
sequence which corresponds to the location of an intron-exon junction in the functional gene. A 4 bp duplication that occurs at amino acid residue 683 of the wild-type coding sequence results in a frameshift mutation and introduces a premature translational termination codon at this point. This is concordant with the inability to detect a human liver transcript corresponding to the
pseudogene
by
nuclease S1
mapping analysis. The 3' end of the
pseudogene
is characterized by a 62 bp segment composed mainly of repeated TC dinucleotides. On the basis of genomic Southern blot analysis performed under high-stringency conditions, the
pseudogene
that we have identified seems to comprise the only sequence in the human genome that is closely related to the wild-type gene. Using somatic cell hybridization, we have mapped the
pseudogene
to human chromosome 8.(ABSTRACT TRUNCATED AT 250 WORDS)
...
PMID:Isolation and characterization of a processed gene for human ceruloplasmin. 342 2
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