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At least 613 records · Page 34Linked to original sources
PubMed1997

Apolipoprotein A-1 of Japanese quail: cDNA sequence and modulation of tissue expression by cholesterol feeding.

Apolipoprotein (apo) A-1 cDNA was amplified by the reverse-transcriptase-polymerase chain reaction (RT-PCR). Primers were synthesized according to the nucleotide sequence of chicken apo A-1, and the identity of apo A-1 cDNA was confirmed by comparing with the N-terminal amino acid sequence. The open reading frame of apo A-1 cDNA consists of 795 nucleotides, and it is capable of coding a polypeptide of 264 amino acids. A comparison between quail and chicken apo A-1 revealed 94.5% homology in the nucleotide sequence and 91.7% homology in the amino acid sequence. There was a similar 11- or 22-amino acid repeat in quail apo A-1 as was the case for chicken apo A-1. Apo A-1 mRNA was evaluated to be 1.4 k in length and was expressed in various tissues of Japanese quail: the liver, small intestine, lung, kidney, heart, and muscle. A quantitative evaluation, however, revealed that the liver and small intestine were the major organs for apo A-1 synthesis, accounting for more than 90% of the total expression of apo A-1 mRNA. Besides apo A-1 mRNA (1.4 k in length), a transcript of 4.1 k was detected in all the tissues examined, with a magnitude ranging from 5 to 10% of the apo A-1 mRNA level. The effect of cholesterol level on the expression of apo A-1 mRNA was studied to address the physiological significance of apo A-1 in the liver, small intestine, and muscle. The level of cholesterol in the liver and breast muscle was increased by feeding with cholesterol and reached a saturation level at day 7. There was also a temporal rise of cholesterol level at day 7 in the small intestine. Dietary cholesterol increased the expression of apo A-1 mRNA two fold in both the liver and small intestine. This was not the case for breast muscle, in which the expression of apo A-1 mRNA was not modulated by the cholesterol level.

Amino Acid Sequence↗
PubMed2000

Molecular cloning and complete cDNA sequences of the ribosomal proteins rpl34 and rpl44 from Aedes triseriatus mosquitoes.

We present the complete cDNA and deduced amino acid sequences of the 60S ribosomal subunit proteins, rpL34 and rpL44, from Aedes triseriatus mosquitoes. The rpL34 cDNA is 554 nucleotides in length and encodes a 139 amino acid protein with a calculated molecular mass of 15 732 daltons. The putative protein displays strong sequence similarity to rpL34 of Aedes albopictus mosquitoes (92%), humans (60%) and rats (58%). The protein is highly basic and contains a C-terminal repetitive-alanine domain and four putative nucleolar localization signals. The rpL44 cDNA consists of 450 nucleotides and encodes a 104 amino acid protein with a calculated molecular mass of 12 544 daltons. The putative protein displays strong sequence similarity to rpL44 of Brugia malayi (87%), Caenorhabditis elegans (86%) and humans (85%). The protein is highly basic and contains a putative nucleolar localization signal. The mRNAs for both rpL34 and rpL44 were detected in biosynthetically active (embryonating) and dormant (diapausing) Ae. triseriatus embryos by RT-PCR analysis.

Aedes↗
PubMed1991

The cDNA sequence and primary structure of the chicken transferrin receptor.

Recombinant cDNA clones encoding the chicken transferrin receptor (cTR) have been isolated and sequenced. Comparison of the deduced primary structure of cTR with those of the human transferrin receptor (hTR) and mouse transferrin receptor (mTR) shows that their size, hydropathy profile, location of sites for posttranslational modifications, and domain organization are highly similar. The cytoplasmic domain of cTR contains the motif Tyr-Xaa-Arg-Phe (YXRF) that is the recognition signal for high-efficiency endocytosis of hTR. The cTR has several highly conserved regions within its extracellular domain, including those flanking the putative N-glycosylation sites. Overall, however, the extracellular domain of cTR is only 53% identical to the extracellular domains of hTR and mTR. The cTR also lacks three of the six Cys residues found in the extracellular domains of the mammalian TRs. These differences can account for functional and structural properties that distinguish cTR and mammalian TRs.

Amino Acid Sequence↗
PubMed1993

The human Ig-beta cDNA sequence, a homologue of murine B29, is identical in B cell and plasma cell lines producing all the human Ig isotypes.

The B cell Ag receptor complex consists of at least two disulfide-linked, heterodimeric structures: the clonally restricted membrane Ig (mIg) molecule and the nonpolymorphic Ig-alpha:Ig-beta protein dimer. The latter molecule is encoded by two separate genes, mb-1 and B29. The DNA sequences of murine and human mb-1 and murine B29 have been determined previously. This study describes the sequence of the full-length human cDNA homologue of the murine Ig-beta/B29 message. The human sequence codes for a protein that displays the typical subunit features of a transmembrane member of the Ig superfamily. The transmembrane and intracytoplasmic domains exhibit striking nucleotide and amino acid sequence similarity between the two species. These regions show almost complete conservation of areas presumed to be involved in noncovalent interactions with other members of the receptor complex and with intracellular kinases and cytoskeletal components. The only sequence dissimilarity seen in these presumed critical areas involves the Y-E-G-L-N motif, a potential target for tyrosine phosphorylation. In contrast, the extracellular portion is much more divergent. Inasmuch as similar patterns of species diversity have been reported for Ig-alpha, the Ig-alpha and Ig-beta molecules may have coevolved to maintain species-specific extracellular interactions between one another and with mIg. Similar to the Ig-alpha molecule, the Ig-beta sequence is identical in B lineage cells expressing all five Ig isotypes. However, in contrast to the Ig-alpha molecule, the Ig-beta sequence is expressed at apparently similar levels in terminally differentiated, mIg- plasma cells as well as in mIg+, mature B cells. These data suggest that Ig-beta has functions in addition to those associated with surface mIg expression.

Amino Acid Sequence↗
PubMed2001

Rice bicoid-related cDNA sequence and its expression during early embryogenesis.

Bicoid is one of the important Drosophila maternal genes involved in the control of embryo polarity and larvae segmentation. To clone and characterize the rice bicoid-related genes, one cDNA clone, Rb24 (EMBL accession number: AJ2771380), was isolated by screening of rice unmature seed cDNA library. Sequence analysis indicates that Rb24 contains a putative amino acid sequence, which is homologous to unique 8 amino acids sequence within Drosophila bicoid homeodomain (50% identity, 75% similarity) and involves a lys-9 in putative helix 3. Northern blot analysis of rice RNA has shown that this sequence is expressed in a tissue-specific manner. The transcript was detected strongly in young panicles, but less in young leaves and roots. This results are further confirmed with paraffin section in situ hybridization. The signal is intensive in rice globular embryo and located at the apical tip of the embryo, then, along with the development of embryo, the signal is getting reduced and transfers into both sides of embryo. The existence of bicoid-related sequence in rice embryo and the similarity of polar distribution of bicoid and Rb24 mRNA in early embryo development may implicates a conserved maternal regulation mechanism of body axis presents in Drosophila and in rice.

Base Sequence↗
PubMed1998

A partial cDNA sequence of the ovine insulin receptor gene: evidence for alternative splicing of an exon 11 region and for tissue-specific regulation of receptor isoform expression in sheep muscle, adipose tissue and liver.

Insulin is as integral and important to the management of metabolism in ruminants as it is in non-ruminants. The suggestion of a lowered ruminant sensitivity and/or responsivity to insulin may relate more to the insulin receptor than to the hormone itself. We screened an ovine cDNA library using degenerate primers and polymerase chain reaction (PCR) to detect and sequence a cDNA portion corresponding to exons 10, 11 and 12 of the human insulin receptor gene in which a 36 base pair (bp) segment (exon 11) is alternatively spliced to produce two distinct receptor isoforms differing in functional characteristics including binding affinity for insulin. The ovine cDNA segment (nucleotides 671 to 770) displayed 84, 84, and 78% nucleotide homology to equivalent segments from the human, rhesus monkey and rat respectively. Reverse transcription PCR (RT-PCR) of selected tissues (liver, m. longissimus dorsi, m. rectus capitis and omental, perirenal and subcutaneous fats) taken at slaughter from three male, pure Dutch Texel lambs (experiment 1) and five male Texel-Greyface crossbred lambs (experiment 2) revealed two mRNA products in each tissue (including spleen; experiment 2 only) corresponding to cDNAs of molecular sizes 161 and 197 bp -- a difference of 36 bp. Sequence alignment showed the 36 bp segment to be homologous to the alternatively spliced exon 11 region of the human insulin receptor gene and to be highly conserved with that from other species. The abundance of the exon 11(+) isoform in the purebred Texel genotype was significantly higher in liver than in perirenal fat and rectus capitis and longissimus dorsi skeletal muscles (P<0.05) and higher also than in subcutaneous and omental fats (P<0.01). There was, however, no difference in the abundance of the exon 11(+) isoform between the individual muscle and fat depots in this sheep genotype. The abundance of the exon 11(+) isoform in the crossbred Texel genotype was significantly higher in liver (P<0. 05) than in the muscles (rectus capitis, P<0.05; longissimus dorsi, P<0.001), all three fats (P<0.001) and spleen (P<0.001). In the crossbred genotype, the abundance of the exon 11(+) isoform was higher in skeletal muscle than in all three fat depots (P<0.001), in which the isoform abundance was similar. Altered ratios of expression of the two products of this alternative splicing event could determine tissue sensitivity and/or responsivity to insulin and provide a mechanism for the management of nutrient partitioning and nutrient utilisation between tissues which is fundamental to the growth of tissues and manipulation of carcass characteristics in meat-producing animals.

Adipose Tissue↗
PubMed1984

Amino acid sequence of the carboxy-terminal part of an acidic type I cytokeratin of molecular weight 51 000 from Xenopus laevis epidermis as predicted from the cDNA sequence.

The DNA sequence of a clone from a cDNA library made from Xenopus laevis skin is described. This sequence represents the 3'-terminal end of an mRNA which codes for an epidermal cytokeratin polypeptide of mol. wt. 51 000 of the acidic (type I) subfamily as identified by hybridization-selection of mRNAs, followed by gel electrophoretic identification of the polypeptides synthesized by translation in vitro. The partial amino acid sequence of the amphibian cytokeratin shows strong similarity to type I cytoskeletal keratins from human (mol. wt. 50 000) and murine (mol. wt. 59 000) epidermis. In the non alpha-helical tail region the human and the non-mammalian (Xenopus) keratins are more similar to each other than to the murine protein, indicating that the former are equivalent cytokeratin polypeptides and belonging to a special subclass of type I keratin polypeptides devoid of glycine-rich regions in the carboxy-terminal portion. The evolutionary conservativity of the genes coding for cytokeratins is discussed.

Amino Acid Sequence↗
PubMed1984

Human T-cell growth factor: partial amino acid sequence, cDNA cloning, and organization and expression in normal and leukemic cells.

The partial amino acid sequences of human T-cell growth factors (TCGFs) isolated from normal peripheral blood lymphocytes and from a leukemia T-cell line (Jurkat) show that the amino-terminal sequences of the two proteins (15 residues) are identical. Oligonucleotides based on the published Jurkat TCGF DNA sequence were used to isolate six cDNA clones of TCGF mRNA from normal lymphocytes. The predicted amino acid sequence of normal lymphocyte TCGF was identical to the sequence of the Jurkat protein, showing that the differences in biochemical properties of the two proteins result from post-translational events. Amino acid and nucleotide sequence data suggest that TCGF is derived from a precursor polypeptide that is cleaved at the amino terminus but not at the carboxyl terminus. Hybridization of the cloned lymphocyte TCGF cDNA to cellular DNA and RNA strongly suggested that the TCGF gene is expressed as a single mRNA species from a single-copy gene. No differences in the organization of the TCGF gene in normal, leukemic, and human T-cell leukemia/lymphoma virus-infected cells was detected regardless of whether they produce TCGF or not.

Amino Acid Sequence↗
PubMed1987

cDNA sequence of a novel sex-limited protein (Slp) from mice constitutive for Slp expression. Sequence comparisons suggest that Slp has no functional role.

Murine sex-limited protein (Slp) is a serum protein that shares 95% sequence identity with murine complement component C4 but does not have C4 activity. Mouse strain B10.WR, which carries the H-2w7 haplotype, has up to 4 Slp genes and is unusual in that both males and females express Slp. Here we report the sequence of a complete pro-Slp cDNA from this strain that we designate Slpw7.2. We find that the Slpw7.2 sequence differs at multiple dispersed sites from three previously reported Slp sequences: two complete pro-Slp sequences, Slpw7.1 and SlpFM, from the B10.WR and FM strains, respectively, and a partial sequence from the B10.WR strain that is distinct from Slpw7.1 as well. A detailed comparison of the complete Slpw7.1, Slpw7.2, and SlpFM sequences reveals that nucleotide changes that alter the amino acid sequence (replacement substitutions) are accumulating at the same relative rate as changes that do not affect the amino acid sequence (silent substitutions); in addition, the amino acid changes themselves tend to be nonconservative. Our results suggest that at least three Slp genes are transcriptionally active in B10.WR mice; that the protein product of the Slpw7.2 transcript predominates in B10.WR serum; and that the Slp protein probably has no function. The Slp system may be particularly suitable for the study of the evolution in the absence of selective pressures of a gene that encodes a stable protein.

Amino Acid Sequence↗
PubMed1995

cDNA sequence and chromosomal localization of a novel human protein, RBQ-1 (RBBP6), that binds to the retinoblastoma gene product.

We have previously isolated cDNA of a novel protein (RBQ-1, HGMW-approved symbol RBBP6) that binds to the retinoblastoma gene product (pRB). Total nucleotide sequence of the cDNA has now been determined. It encoded a protein of 140 kDa that consists of 948 amino acids and contains multiple repeated sequences like SRS, YRE, and VPPP. The region used for pRB binding was identified on a small region near the C-terminus. We have mapped this gene to 16p11.2-p12 using polymerase chain reaction analysis on a human-hamster hybrid cell panel and chromosomal fluorescence in situ hybridization.

Amino Acid Sequence↗
PubMed1996

The cDNA sequence encoding bovine SP-22, a new defence system against reactive oxygen species in mitochondria.

We have isolated cDNA clones coding SP-22, an antioxidant protein in mitochondria, from a bovine adrenal medulla cDNA library constructed with (lambda)gt11. The largest clone contained the entire coding sequence for mature SP-22. Since the isolated cDNA clones lacked 5'- and 3'-ends, we determined the sequences of both ends by the "Rapid Amplification of cDNA Ends (RACE)" tecnique. The deduced amino acid sequence of the mature protein region was the same as that determined by protein sequencing. Since SP-22 had a mitochondrial targetting signal, its mitochondrial localization was confirmed.

Adrenal Medulla↗
PubMed1993

The cDNA sequence encoding the major intrinsic protein of frog lens.

A cDNA clone encoding the frog lens major intrinsic protein (MIP) has been isolated and sequenced. The predicted protein of 28 kDa has high sequence identity and similarity to mammalian and avian lens MIP sequences. Frog lens MIP is encoded by a transcript of 4.4 kb.

Amino Acid Sequence↗
PubMed1999

HUGE: a database for human large proteins identified by Kazusa cDNA sequencing project.

HUGE is a database for human large proteins newly identified by Kazusa cDNA project, which aims to predict protein primary structures from sequences of human large cDNAs (>4 kb). In particular, cDNA clones capable of coding for large proteins (>50 kDa) are current targets of the project. More than 700 sequences of human cDNAs (average size, 5.1 kb) have been determined to date and deposited in the public databases. Notable information implied from the cDNAs and the predicted protein sequences can be obtained through HUGE via the World Wide Web at URL http://www.kazusa.or.jp/huge

Cloning, Molecular↗
PubMed1990

Primary structure of human plasma glutathione peroxidase deduced from cDNA sequences.

Human plasma glutathione peroxidase (GSHPx) has been shown to be a selenium-containing enzyme immunologically distinct from cellular GSHPx. Oligonucleotide probes, based on the partial amino acid sequence of plasma GSHPx, were synthesized and used to screen a human placenta cDNA library. Nucleotide sequence analysis of the obtained clones revealed that GSHPx consisted of a 678-base pair open reading frame coding for a 226-amino acid polypeptide with a Mr of 25,389. About 50% of the deduced amino acid sequence was confirmed by partial amino acid sequencing of the peptides in a lysine endopeptidase-digest of the purified enzyme. The amino acid sequence exhibited only 44% homology with that of human cellular GSHPx. Northern blot analysis revealed a single transcript of 2.2 kilobases in the poly(A)+ RNA fractions of human placenta and HepG2 (a human hepatic cell line), but not that of human liver and endothelial cells.

Amino Acid Sequence↗
PubMed1996

Simple strategy for sequencing cDNA clones.

We describe a simple method for constructing subclones containing overlapping nested deletions from cDNA clones (both lambda phage clones and plasmid clones). A PCR-amplified insert is partially digested with 4-cutter restriction enzyme(s). Complete digestion of this DNA with two restriction enzymes, having unique cutting sites at one or the other end of the amplified DNA, creates two sets of overlapping nested subfragments. When recloned into each of two doubly cut pBluescript plasmid vectors, only the two sets of nested subfragments are produced. Minimal nested sets can be constructed by screening subclones using colony PCR, and this set can then be used to determine the entire sequence of the cDNA clone. This method requires only a single cloning step and can be generated from an insert that is amplified directly from a lambda phage clone. This procedure eliminates the sequencing redundancy problem inherent in shotgun cloning, allows large clones to be sequenced using universal primers only and is well-suited for automated DNA-sequencing. Using this method, we successfully sequenced five cDNA clones of five Drosophila subobscura genes.

Animals↗
PubMed1986

Chromosomal protein HMG-17. Complete human cDNA sequence and evidence for a multigene family.

Antibodies elicited against chromosomal protein HMG-17, purified from calf, were used to screen a human lambda gt11 cDNA expression library and isolate the full length cDNA coding for this protein. Sequence analysis reveals that the nucleotide distribution along this cDNA is highly asymmetric. The amino acid sequence, deduced from the reading frame, reveals that the human HMG-17 is, respectively, 96 and 92% homologous with the calf and chicken protein. The amino acid substitution are conservative suggesting evolutionary constraints on the conformation of the protein. The human genome contains 35-50 HMG-17 gene copies which, as revealed by Southern analysis, are distributed at several loci. Northern analysis of total RNA isolated from 3 human cell lines, indicates that each cell contains a single-size mRNA coding for this protein. Nucleotide sequences which cross-hybridize, under stringent conditions, with the human HMG-17 cDNA are present in the genome of rodents and absent from the genomes of sea urchin, Drosophila, and yeast. The availability of a probe for the HMG-17 gene may help elucidate the cellular role of this protein which may confer specific conformations to transcribable regions in the genome.

Amino Acid Sequence↗
PubMed1994

A complete cDNA sequence for core I protein subunit of human ubiquinol-cytochrome c reductase.

Core I protein is the largest subunit of ubiquinol-cytochrome c reductase. We have isolated a complete cDNA clone of 1575 bp encoding the precursor to this protein by screening a human fibroblast cDNA library. Nucleotide sequence comparison showed that the human core I protein cDNA is about 85% homologous with the reported bovine counter part but with a large difference in the length of coding region which consists of 480 amino acids in human and 362 amino acids in bovine. Human core I protein is presumed to contain a presequence of 34 amino acids. Amino acid sequence alignment study showed that the predicted human core I protein has a significant homology with other members of matrix processing peptidase (MPP) and processing enhancing protein (PEP) family.

Amino Acid Sequence↗
PubMed1990

Complete cDNA sequence and chromosomal localization of mouse alpha 1-antitrypsin.

A cDNA encoding the complete open reading frame of murine alpha 1-antitrypsin has been cloned and sequenced. The nucleic acid and predicted amino acid sequences show homology to human alpha 1-antitrypsin and demonstrate the preservation of critical structural determinants for intracellular targeting, carbohydrate attachment, and catalytic function. The alpha 1-antitrypsin gene locus (Aat) has been localized on murine chromosome 12E----F by in situ hybridization.

Amino Acid Sequence↗