scholarly journals The Australian brushtail possum (Trichosurus vulpecula) gonadotrophin alpha-subunit: analysis of cDNA sequence and pattern of expression

1998 ◽  
Vol 20 (3) ◽  
pp. 345-353 ◽  
Author(s):  
AE Fidler ◽  
SB Lawrence ◽  
DM Vanmontfort ◽  
DJ Tisdall ◽  
KP McNatty
Keyword(s):  
Cdna Sequence ◽  
Amino Acid Sequences ◽  
Alpha Subunit ◽  
Brushtail Possum ◽  
Pituitary Cells ◽  
Trichosurus Vulpecula ◽  
Coding Region ◽  
Recognition Sequence ◽  
Subunit Protein ◽  
Evolutionarily Conserved

A cDNA sequence from the gonadotrophin alpha-subunit mRNA of Australian brushtail possum (Trichosurus vulpecula) has been determined and analysed. Comparison with seven eutherian mammalian gonadotrophin alpha-subunit gene sequences revealed an average of 82.6% homology between the coding region nucleotide sequences and 88.8% identity between the predicted amino acid sequences. The predicted possum gonadotrophin alpha-subunit protein has ten evolutionarily conserved cysteine residues, two potential N-linked glycosylation sites and a putative enzyme recognition sequence which it has been suggested is required for sulphation of carbohydrate moieties. Comparison of the possum gonadotrophin alpha-subunit 3' untranslated region (UTR) sequence with the 3' UTRs of eutherian alpha-subunit transcripts revealed sequence homology. In particular, an 18 nucleotide imperfect palindromic sequence present in the possum 3' UTR, with the potential to form a hairpin loop, was found to be evolutionarily conserved and present in five out of seven eutherian alpha-subunit 3' UTR sequences. In situ hybridization localized the transcripts to a sub-population of anterior pituitary cells presumed to be gonadotrophs and thyrotrophs. In summary, these results indicate considerable conservation of the structure and function of the gonadotrophin alpha-subunit protein since the divergence of the marsupial and eutherian mammalian lineages.

scholarly journals cDNA sequence analysis, gene expression and protein localisation of the inhibin alpha-subunit of Australian brushtail possum (Trichosurus vulpecula)

1998 ◽  
Vol 21 (2) ◽  
pp. 141-152 ◽  
Author(s):  
D Vanmontfort ◽  
AE Fidler ◽  
DA Heath ◽  
SB Lawrence ◽  
DJ Tisdall ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cdna Sequence ◽  
In Situ Hybridisation ◽  
Precursor Protein ◽  
Alpha Subunit ◽  
Brushtail Possum ◽  
Trichosurus Vulpecula ◽  
Follicular Growth ◽  
Reading Frame ◽  
Stage Of Development

An inhibin alpha-subunit cDNA sequence from the Australian brushtail possum (Trichosurus vulpecula) has been identified and analysed. The cDNA includes an open reading frame encoding a predicted precursor protein of 361 amino acids. The predicted protein sequence includes four possible proteolytic cleavage sites, 12 evolutionarily conserved cysteine residues and three potential N-linked glycosylation sites. The mature alpha-subunit is the carboxyl terminal fragment (alphaC) consisting of 131 amino acids. The full-length precursor protein shows a mean identity with eutherian homologues of 69.8%. The homology is not evenly distributed, with the putative alphaC fragment showing the highest level (79.7%). Using Northern hybridisation, an alpha-subunit transcript of approximately 1.6 kb was detected in adult possum ovary. Using in situ hybridisation and immunocytochemistry, inhibin alpha-subunit was localised exclusively to the granulosa cell layers of follicles. Hybridisation and immunostaining for the inhibin alpha-subunit were first observed in granulosa cells of primary follicles and the expression continued throughout all stages of follicular growth. Inhibin alpha-subunit mRNA and protein were also detected in cells of the corpus luteum. In summary, results indicate considerable conservation of the structure and possible function of the inhibin alpha-subunit protein since the divergence of the marsupial and eutherian mammalian lineages. The expression data suggest that, in the adult possum, inhibin may have a role in ovarian follicular growth from the primary stage of development.

scholarly journals Bovine cytosolic IMP/GMP-specific 5′-nucleotidase: cloning and expression of active enzyme in Escherichia coli

1997 ◽  
Vol 328 (2) ◽  
pp. 483-487 ◽  
Author(s):  
Simone ALLEGRINI ◽  
Rossana PESI ◽  
Maria Grazia TOZZI ◽  
J. Carol FIOL ◽  
B. Robert JOHNSON ◽  
...  
Keyword(s):  
Cdna Sequence ◽  
Amino Acid Sequences ◽  
Cloning And Expression ◽  
Kinetic Properties ◽  
Coding Region ◽  
Northern Blots ◽  
Sds Page ◽  
Calf Thymus ◽  
Rapid Amplification ◽  
First Time

A cDNA coding for bovine cytosolic IMP/GMP-specific 5ʹ-nucleotidase endowed with phosphotransferase activity was cloned from calf thymus RNA, by 5ʹ and 3ʹ rapid amplification of cDNA ends protocols (5ʹ and 3ʹ RACE). Two products were isolated: a 5ʹ RACE 1.6 kb fragment and a 3ʹ RACE 2.0 kb fragment, with an overlapping region of 505 bp, leading to a total length of approx. 2951 bp. The similarity in the coding region to that of the human 5ʹ-nucleotidase cDNA sequence [Oka, Matsumoto, Hosokawa and Inoue (1994) Biochem. Biophys. Res. Commun. 205, 917-922], indirectly identified as a 5ʹ-nucleotidase, was 94% and the deduced amino acid sequences were 99.5% identical. The bovine cDNA sequence included the sequences codifying for six peptides obtained from 5ʹ-nucleotidase/phosphotransferase purified from calf thymus. Northern blots of human mRNA species from different tissues showed a 3.6 kb mRNA expressed at equal levels in most tissues. The cDNA was cloned into a pET-28c expression vector and the protein obtained after induction had a molecular mass of 61 kDa under SDS/PAGE. It exhibited both 5ʹ-nucleotidase and phosphotransferase activity, as well as immunological and kinetic properties similar to those of the enzyme purified from calf thymus. This is the first time that a fully active recombinant 5ʹnucleotidase has been described.

The follicle-stimulating hormone β-subunit gene of the common brushtail possum (Trichosurus vulpecula): analysis of cDNA sequence and expression

1997 ◽  
Vol 9 (8) ◽  
pp. 795 ◽  
Author(s):  
Stephen B. Lawrence ◽  
Dominique M. Vanmontfort ◽  
David J. Tisdall ◽  
Kenneth P. McNatty ◽  
Andrew E. Fidler
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Cdna Sequence ◽  
Follicle Stimulating Hormone ◽  
Evolutionary Conservation ◽  
Brushtail Possum ◽  
Predicted Amino Acid Sequence ◽  
Trichosurus Vulpecula ◽  
Β Subunit ◽  
Subunit Gene

Reverse transcription–PCR has been used to obtain a cDNA sequence from the follicle-stimulating hormone (FSH) β-subunit gene of the Australian brushtail possum (Trichosurus vulpecula). Comparisons of the possum FSHβ-mRNA coding region nucleotide sequence with that of six eutherian mammal homologues reveals a mean percent identity of 77·3% and 76·8% at the nucleotide and predicted amino acid-sequence levels respectively. Furthermore, the predicted amino acid sequence of the possum FSHβ mature protein shows evolutionary conservation of twelve cysteine residues and two potential N-linked glycosylation sites. The protein lacks the CAGY motif present in most reported glycoprotein β-subunit sequences. The translation termination codon and consensus polyadenylation sequence overlap, a feature observed in other mammalian FSHβ genes. Northern hybridization of total RNA from adult female possum pituitary revealed three hybridizing transcripts of approximately 2·8, 1·2 and 0·5 kb which may arise from utilizing alternative polyadenylation signals. In situ hybridization localized the FSHβ transcripts to a sub-population of anterior pituitary cells interpreted as being gonadotropes. In summary the results indicate considerable evolutionary conservation of the structure of the FSH b-subunit gene between the marsupial and eutherian mammalian lineages.

scholarly journals Murine erythropoietin gene: cloning, expression, and human gene homology.

1986 ◽  
Vol 6 (3) ◽  
pp. 849-858 ◽  
Author(s):  
C B Shoemaker ◽  
L D Mitsock
Keyword(s):  
Amino Acid ◽  
Genomic Library ◽  
Cdna Probe ◽  
Amino Acid Sequences ◽  
Nucleotide Sequence Analysis ◽  
Coding Region ◽  
Transcription Control ◽  
Erythroid Progenitor ◽  
Human Genes ◽  
Cell Expression

The gene for murine erythropoietin (EPO) was isolated from a mouse genomic library with a human EPO cDNA probe. Nucleotide sequence analysis permitted the identification of the murine EPO coding sequence and the prediction of the encoded amino acid sequence based on sequence conservation between the mouse and human EPO genes. Both the coding DNA and the amino acid sequences were 80% conserved between the two species. Transformation of COS-1 cells with a mammalian cell expression vector containing the murine EPO coding region resulted in secretion of murine EPO with biological activity on both murine and human erythroid progenitor cells. The transcription start site for the murine EPO gene in kidneys was determined. This permitted tentative identification of the transcription control region. The region included 140 base pairs upstream of the cap site which was over 90% conserved between the murine and human genes. Surprisingly, the first intron and much of the 5'- and 3'-untranslated sequences were also substantially conserved between the genes of the two species.

scholarly journals Preovulatory follicle development and ovulation in the brushtail possum (Trichosurus vulpecula) monitored by repeated laparoscopy

Reproduction ◽  
1997 ◽  
Vol 110 (2) ◽  
pp. 361-370 ◽  
Author(s):  
J. L. Crawford ◽  
G. H. Shackell ◽  
E. G. Thompson ◽  
B. J. McLeod ◽  
P. R. Hurst
Keyword(s):  
Brushtail Possum ◽  
Follicle Development ◽  
Trichosurus Vulpecula ◽  
Preovulatory Follicle
Sign in / Sign up

Export Citation Format

Share Document