scholarly journals Oestrogen Sulfotransferase: Molecular Cloning and Sequencing of cDNA for the Bovine Placental Enzyme

1988 ◽  
Vol 41 (4) ◽  
pp. 507 ◽  
Author(s):  
Allan R Nash ◽  
Wendy K Glenn ◽  
Stephen S Moore ◽  
Judith Kerr ◽  
Adrienne R Thompson ◽  
...  
Keyword(s):  
Transcriptional Control ◽  
Sequence Data ◽  
Apparent Molecular Weight ◽  
Peptide Sequence ◽  
Binding Region ◽  
Cloning And Sequencing ◽  
Reading Frame ◽  
Physico Chemical ◽  
Malignant State ◽  
Steroid Binding

The female sex hormone, oestrogen, plays a central role in breast cell proliferation in both the normal and malignant state. It controls transcription from several genes, including that for the progesterone receptor, and in endometrial tissue, via this receptor, it controls the gene for the enzyme oestrogen sulfotransferase. This enzyme may control the level of the oestrogen receptor by sulfurylating free oestradiol. To study the mode of transcriptional control exercised by oestrogen, bovine oestrogen sulfotransferase cDNA has been cloned and the nucleotide sequence determined. The message, of which 1812 bases have been sequenced, contains an open reading frame of 885 bases which encode a protein of 295 amino acids and a maximum apparent molecular weight of 34 600. The deduced protein sequence is supported by existing peptide sequence data and appears to contain a steroid-binding region. Some physico-chemical characteristics of the enzyme appear to differ markedly from those previously reported.

scholarly journals Cloning of cDNA for the γ-subunit of mammalian translation initiation factor 2B, the guanine nucleotide-exchange factor for eukaryotic initiation factor 2

1996 ◽  
Vol 318 (2) ◽  
pp. 631-636 ◽  
Author(s):  
Nigel T PRICE ◽  
Scot R. KIMBALL ◽  
Leonard S. JEFFERSON ◽  
Christopher G. PROUD
Keyword(s):  
Sequence Data ◽  
Pcr Primers ◽  
Genome Project ◽  
Translation Initiation Factor ◽  
Peptide Sequence ◽  
Initiation Factor ◽  
Nucleotide Exchange ◽  
Reading Frame ◽  
C Elegans ◽  
Γ Subunit

Peptide sequence data were obtained from rabbit protein synthesis initiation factor subunit eIF2Bγ. Searching the database of expressed sequence tags (dbEST) revealed nucleotide sequences potentially encoding human eIF2Bγ that contained peptides corresponding to those from the rabbit subunit. PCR primers were derived from these sequences and used to generate a probe. This was used to screen a rat skeletal muscle cDNA library, and a clone encoding rat eIF2Bγ was isolated. This cDNA gave a product in coupled transcription/translation that co-migrated with the γ-subunit of purified eIF2B under SDS/PAGE. The sequence of this rat eIF2Bγ cDNA is reported. The protein sequence shows homology with that of yeast eIF2Bγ (the GCD1 gene product). We have also identified an open reading frame from the Caenorhabditis elegans genome project that probably encodes the γ-subunit of C. elegans eIF2B. All these sequences show similarity to nucleotidyl- and acyltransferases, as previously reported for GCD1 [Koonin (1995) Protein Sci. 4, 1608–1617], and contain conserved motifs potentially involved in nucleotide binding. They also contain ‘I-patch’ motifs: isoleucine-rich hexamer repeats that have been associated with the binding of acyl groups in bacterial acyltransferases. The roles of these motifs are discussed in relation to the known properties of eIF2B.

Physico-Chemical Properties of Recombinant Desulphatohirudin

1990 ◽  
Vol 63 (03) ◽  
pp. 499-504 ◽  
Author(s):  
A Electricwala ◽  
L Irons ◽  
R Wait ◽  
R J G Carr ◽  
R J Ling ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Gel Filtration ◽  
Chemical Properties ◽  
Alpha Helix ◽  
Apparent Molecular Weight ◽  
Correlation Spectroscopy ◽  
Nmr Studies ◽  
Recombinant Hirudin ◽  
Physico Chemical ◽  
Recombinant Molecule

SummaryPhysico-chemical properties of recombinant desulphatohirudin expressed in yeast (CIBA GEIGY code No. CGP 39393) were reinvestigated. As previously reported for natural hirudin, the recombinant molecule exhibited abnormal behaviour by gel filtration with an apparent molecular weight greater than that based on the primary structure. However, molecular weight estimation by SDS gel electrophoresis, FAB-mass spectrometry and Photon Correlation Spectroscopy were in agreement with the theoretical molecular weight, with little suggestion of dimer or aggregate formation. Circular dichroism studies of the recombinant molecule show similar spectra at different pH values but are markedly different from that reported by Konno et al. (13) for a natural hirudin-variant. Our CD studies indicate the presence of about 60% beta sheet and the absence of alpha helix in the secondary structure of recombinant hirudin, in agreement with the conformation determined by NMR studies (17)

Isolation of a cDNA corresponding to a developmentally regulated transcript in rat intestine

1988 ◽  
Vol 8 (10) ◽  
pp. 4243-4249
Author(s):  
J Filmus ◽  
J G Church ◽  
R N Buick
Keyword(s):  
Cell Line ◽  
Sequence Data ◽  
Human Colon ◽  
Intestinal Cell ◽  
Homologous Gene ◽  
Rat Intestine ◽  
Human Colon Carcinoma Cell ◽  
Reading Frame ◽  
Intestinal Cell Line ◽  
High Level

We report the isolation of a cDNA clone corresponding to a transcript that is accumulated differentially in rat intestine during development. Clone OCI-5 was selected from the rat intestinal cell line IEC-18, which represents primitive intestinal epithelial crypt cells. Expression was high in rat fetal intestine between 15 and 19 days of development and thereafter was progressively down regulated, becoming undetectable after weaning. Clone OCI-5 detected homologous sequences in human and murine cells. In particular, a high level of expression was detected in CaCo-2, a human colon carcinoma cell line, which is known to express molecules characteristic of fetal small intestinal cells. Expression of a homologous gene was also detected in F9 murine teratocarcinoma cells when they were induced to differentiate into parietal or visceral endodermlike cells. When IEC-18 cells were transformed by activated H-ras or v-src genes, expression of clone OCI-5 was suppressed; the degree of down-regulation correlated with the extent of morphological change induced in the transformed IEC-18 cells. The sequence of clone OCI-5 showed an open reading frame that was capable of encoding a protein of 597 amino acids, but no strong homology was found with any of the proteins registered in the protein sequence data base.

Molecular cloning and sequence analysis of putative chicken prolactin cDNA

1989 ◽  
Vol 2 (1) ◽  
pp. 21-30 ◽  
Author(s):  
M.C. Hanks ◽  
J.A. Alonzi ◽  
P.J. Sharp ◽  
H.M. Sang
Keyword(s):  
Amino Acids ◽  
Genomic Dna ◽  
Signal Sequence ◽  
Peptide Sequence ◽  
Mrna Levels ◽  
Reading Frame ◽  
Ring Dove ◽  
Prolactin Gene ◽  
Pituitary Glands ◽  
High Degree

ABSTRACT A cDNA library was prepared from mRNA isolated from anterior pituitary glands of incubating bantam hens, in which prolactin mRNA levels were predicted to be very high. Nine clones, representing abundant mRNA species, were identified and shown to contain homologous sequences. Two clones, of 871 bp and 580 bp, were analysed by DNA sequencing. The shorter clone was found to be a truncated cDNA product but otherwise identical to the longer clone. The 871 bp cDNA, PRL101, contains an open reading frame capable of encoding a polypeptide of 229 amino acids. This putative polypeptide has a high degree of homology to mammalian prolactins (approximately 70%), strongly suggesting that PRL101 encodes chicken preprolactin. The protein was predicted to have a 30 amino acid signal sequence which would be cleaved off to give a mature protein of 199 amino acids. The peptide sequence also had a 26% homology to chicken growth hormone, which is related to prolactin. This similarity confirms the conclusion that PRL101 is a chicken prolactin cDNA clone. An abundant mRNA of approximately 880 b was detected in poly(A)+ RNA from pituitary glands probed with PRL101. Analysis of chicken genomic DNA showed that there is one copy of the prolactin gene in the genome. PRL101 hybridized strongly to genomic DNA from closely related galliforms (quail and turkey) and less strongly to DNA from more distantly related species (duck and ring dove).

scholarly journals Novel Genomic Locus with Atypical G+C Content that Is Required for Extracellular Polysaccharide Production and Virulence in Xanthomonas oryzae pv. oryzae

2001 ◽  
Vol 14 (11) ◽  
pp. 1335-1339 ◽  
Author(s):  
Sridhar Dharmapuri ◽  
J. Yashitola ◽  
M. R. Vishnupriya ◽  
Ramesh V. Sonti
Keyword(s):  
Sequence Data ◽  
Extracellular Polysaccharide ◽  
Genomic Region ◽  
Xanthomonas Oryzae ◽  
Genomic Locus ◽  
Reading Frame ◽  
Glycosyl Transferase ◽  
Clone Sequence ◽  
Gum Gene Cluster ◽  
Transposon Insertions

Three exopolysaccharide (EPS)- and virulence-deficient mutants of Xanthomonas oryzae pv. oryzae, the causal agent of bacterial leaf blight of rice, were isolated by Tn5 mutagenesis. These insertions are not located within the gum gene cluster. A 40-kb cosmid clone that restored EPS production and virulence to all three mutants was isolated, and the three transposon insertions were localized to contiguous 4.3- and 3.5-kb EcoRI fragments that are included in this clone. Sequence data indicate that two of the transposon insertions are in genes that encode a putative sugar nucleotide epimerase and a putative glycosyl transferase, respectively; the third insertion is located between the glycosyl transferase gene and a novel open reading frame (ORF). A 5.5-kb genomic region in which these three ORFs are located has a G+C content of 51.7%, quite different from the G+C content of approximately 65.0% that is typical of X. oryzae pv. oryzae. Homologues of this locus have not yet been reported in any other xanthomonad.

Expression patterns of the genes that encode lectin or lectin-related polypeptides in Robinia pseudoacacia

1999 ◽  
Vol 26 (5) ◽  
pp. 495 ◽  
Author(s):  
Kazumasa Yoshida ◽  
Kiyoshi Tazaki
Keyword(s):  
Amino Acid ◽  
Nucleotide Sequence ◽  
Amino Acid Sequence ◽  
Sequence Data ◽  
Expression Patterns ◽  
Robinia Pseudoacacia ◽  
Regulation Of Expression ◽  
Nucleotide Sequence Data ◽  
Reading Frame ◽  
Inner Bark

Three genomic clones (Rplec2, Rplec5 and Rplec6) and a cDNA clone (LECRPA4) that encoded lectin or lectin-related polypeptides were isolated from Robinia pseudoacacia L. A comparison of the nucleotide sequences of Rplec2 and a previously reported cDNA for the subunit indicated that Rplec2 encoded the 29 kDa subunit of the inner-bark lectin RPbAI. Rplec5 encoded a polypeptide whose deduced amino acid sequence was 96.1% identical to that of a subunit of seed lectin. The amino acid sequence deduced from the open reading frame of Rplec6 showed 61.1% identity to that encoded by Rplec5. LECRPA4 was isolated from an inner bark cDNA library and appeared to encode the 26 kDa subunit of inner-bark lectin RPbAII. The expression patterns of the various genes in tissues were examined by the reverse transcriptase-polymerase chain reaction (RT-PCR) with appropriate primers. Rplec2 transcripts were detected in the inner bark and roots. Rplec5 transcripts were detected in the inner bark, seeds and roots. No Rplec6 transcripts were detected in all tissues examined. LECRPA4 transcripts were found in leaves and in the inner bark. The level of expression of Rplec2 in the inner bark appeared to be similar in samples collected in different years and from different trees, whereas levels of expression of Rplec5 and LECRPA4 varied. These results suggest the differential regulation of expression of members of the lectin gene family in tissues of R. pseudoacacia. The nucleotide sequence data reported herein will appear in the DDBJ, EMBL and GenBank Nucleotide Sequence Databases under the accession numbers AB 012632 (Rplec2), AB012633 (Rplec5), AB012634 (Rplec6) and AB012635 (LECRPA4).

scholarly journals coil: an R package for cytochrome c oxidase I (COI) DNA barcode data cleaning, translation, and error evaluation

Genome ◽  
2020 ◽  
Vol 63 (6) ◽  
pp. 291-305 ◽  
Author(s):  
Cameron M. Nugent ◽  
Tyler A. Elliott ◽  
Sujeevan Ratnasingham ◽  
Sarah J. Adamowicz
Keyword(s):  
Cytochrome C ◽  
Cytochrome C Oxidase ◽  
Sequence Data ◽  
Dna Barcode ◽  
R Package ◽  
Data Generation ◽  
Error Identification ◽  
Reading Frame ◽  
Biological Contaminants ◽  
And Function

Biological conclusions based on DNA barcoding and metabarcoding analyses can be strongly influenced by the methods utilized for data generation and curation, leading to varying levels of success in the separation of biological variation from experimental error. The 5′ region of cytochrome c oxidase subunit I (COI-5P) is the most common barcode gene for animals, with conserved structure and function that allows for biologically informed error identification. Here, we present coil ( https://CRAN.R-project.org/package=coil ), an R package for the pre-processing and frameshift error assessment of COI-5P animal barcode and metabarcode sequence data. The package contains functions for placement of barcodes into a common reading frame, accurate translation of sequences to amino acids, and highlighting insertion and deletion errors. The analysis of 10 000 barcode sequences of varying quality demonstrated how coil can place barcode sequences in reading frame and distinguish sequences containing indel errors from error-free sequences with greater than 97.5% accuracy. Package limitations were tested through the analysis of COI-5P sequences from the plant and fungal kingdoms as well as the analysis of potential contaminants: nuclear mitochondrial pseudogenes and Wolbachia COI-5P sequences. Results demonstrated that coil is a strong technical error identification method but is not reliable for detecting all biological contaminants.

A β-Mannanase from Bacillus subtilis B36: Purification, Properties, Sequencing, Gene Cloning and Expression in Escherichia coli

2006 ◽  
Vol 61 (11-12) ◽  
pp. 840-846 ◽  
Author(s):  
Ya Nan Li ◽  
Kun Meng ◽  
Ya Ru Wang ◽  
Bin Yao
Keyword(s):  
Escherichia Coli ◽  
Enzyme Activity ◽  
Bacillus Subtilis ◽  
Specific Activity ◽  
Ph Value ◽  
High Specificity ◽  
Apparent Molecular Weight ◽  
Cloning And Expression ◽  
Gene Cloning And Expression ◽  
Reading Frame

Abstract MANB36, a secrete endo-β-1,4-D-mannanase produced by Bacillus subtilis B36, was puri­fied to homogeneity from a culture supernatant and characterized. The optimum pH value for the mannanase activity of MANB36 is 6.4 and the optimum temperature is 50 °C. The enzyme activity of MANB36 is remarkably thermostable at 60 °C and the specific activity of MANB36 is 927.84 U/mg. Metal cations (except Hg2+ and Ag+), EDTA and 2-mercaptoetha- nol (2-ME) have no effects on enzyme activity. This enzyme exhibits high specificity with the substituted galactomannan locust bean gum (LBG). The gene encoding for MANB36, manB36, was cloned by PCR and sequenced. manB36 contains a single open reading frame (ORF) consisting of 1104 bp that encodes a protein of 367 amino acids. The predicted mo­lecular weight of 38.13 kDa, calculated by the deduced protein of the gene manB36 without signal peptide, coincides with the apparent molecular weight of 38.0 kDa of the purified MANB36 estimated by SDS-PAGE. The mature protein of MANB36 has been expressed in Escherichia coli BL21 and the expressed mannanase has normal bioactivity.

Sign in / Sign up

Export Citation Format

Share Document