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 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.

A novel CYP450 gene (CYP4V18) of the shrimp Penaeus schmitti Burkenroad, 1936 (Decapoda, Dendrobranchiata): cloning and down-regulation of CYP4V18 expression by the imidazole derivative KK-42

Crustaceana ◽  
2013 ◽  
Vol 86 (10) ◽  
pp. 1179-1190
Author(s):  
X. Xichao ◽  
W. Wenfeng ◽  
L. Xin ◽  
W. Xueshen ◽  
L. Li ◽  
...  
Keyword(s):  
Amino Acids ◽  
High Performance ◽  
Cdna Sequence ◽  
Mrna Level ◽  
Open Reading Frame ◽  
Control Group ◽  
Reverse Phase ◽  
Reading Frame ◽  
Cyp4 Family ◽  
Experimental Stage

The CYP4 family is the most ancient and diversified group of P450 and some members are associated with ecdysteroid metabolism in arthropods. In an attempt to research the possible molecular mechanism of KK-42 action on Penaeus schmitti Burkenroad, 1936, one CYP4 cDNA sequence from the hepatopancreas was first cloned and its spatiotemporal expressions were analysed using real-time PCR, and the haemolymph 20-hydroxyecdysone (20E) level was measured by reverse-phase high-performance liquid chromatography. Results showed that the full-length cDNA contains a 1545 bp open reading frame encoding a predicted protein of 515 amino acids named CYP4V18 by the P450 nomenclature committee. The highest cyp4v18 mRNA level was detected in hepatopancreas. Compared with that of the KK-42 treatment group, the hepatopancreas cyp4v18 expression in the control group increased to >72.3%, even 187.7% () on day 6. The haemolymph 20E levels increased slightly in KK-42-treated shrimps compared to that in controls during the experimental stage. The results show that the hepatopancreas CYP4V18 is not involved in 20E biosynthesis, and likely plays other roles in the shrimp, P. schmitti.

Cloning and sequence analysis of a cDNA encoding an androgen-dependent mouse seminal vesicle secretory protein

1993 ◽  
Vol 10 (3) ◽  
pp. 279-288 ◽  
Author(s):  
C Guilbaud ◽  
A M Simon ◽  
G Veyssière ◽  
C Jean
Keyword(s):  
Amino Acids ◽  
Seminal Vesicle ◽  
Cdna Sequence ◽  
Northern Blot Analysis ◽  
Secretory Protein ◽  
Calculated Molecular Mass ◽  
Reading Frame ◽  
The Family ◽  
Sequence Encoding ◽  
High Degree

ABSTRACT We report the cloning and sequencing of a new cDNA sequence encoding a protein from the mouse seminal vesicle. An open reading frame of 297 nucleotides encoded a protein of 99 amino acids with a calculated molecular mass of 11·454 kDa. The first 21 amino acids constituted a signal peptide followed by 78 amino acids encoding the secreted protein. The cDNA sequence comprised a 3′ untranslated region of 226 bp and the polyadenylation signal AATAAA, 19 bp upstream from the poly(A)+ tail. A high degree of homology was found between this protein and members of the family of seminal vesicle secretory (SVS) proteins, especially rat SVS VI. Northern blot analysis indicated the presence of a 0·7 kb mRNA species in the mRNAs of seminal vesicle tissue. Castration resulted in a marked decrease in the level of the 0·7 kb mRNA encoding the protein, whereas administration of testosterone to castrated males restored the 0·7 kb mRNA.

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 Immunogenicity and contraceptive potential of three infertility-relevant zona pellucida 2 epitopes in the marsupial brushtail possum (Trichosurus vulpecula)

Reproduction ◽  
2007 ◽  
Vol 133 (1) ◽  
pp. 177-186 ◽  
Author(s):  
Janine A Duckworth ◽  
Kathryn Wilson ◽  
Xianlan Cui ◽  
Frank C Molinia ◽  
Phil E Cowan
Keyword(s):  
Amino Acids ◽  
Zona Pellucida ◽  
Keyhole Limpet Hemocyanin ◽  
Brushtail Possum ◽  
Trichosurus Vulpecula ◽  
Incomplete Freund’S Adjuvant ◽  
Freund’S Adjuvant ◽  
Freund's Adjuvant ◽  
Phosphate Buffered Saline

In a previous study, three infertility-relevant epitopes of possum ZP2 (Pep12 (amino acids 111–125), Pep31 (amino acids 301–315), and Pep44 (amino acids 431–445)) were identified using sera from possums (Trichosurus vulpecula) immunized with recombinant possum zona pellucida 2 (ZP2) constructs, and a synthetic peptide library of possum ZP2 protein. In this study, the three peptides were conjugated to keyhole limpet hemocyanin and 300 μg of each conjugated peptide were administered subcutaneously to female possums (n = 20 per peptide) in complete Freund’s adjuvant. Immunogen doses were repeated 3 and 6 weeks later using incomplete Freund’s adjuvant. Control animals were immunized with either phosphate-buffered saline only (n = 10) or 300 μg keyhole limpet hemocyanin (n = 10), administered with the same adjuvants. Serum antibodies from animals immunized against these three epitopes bound to the corresponding possum ZP2 peptides, recombinant possum ZP2 protein constructs, and native zona. Possum fertility was assessed following superovulation and artificial insemination. Peptides Pep12 and Pep31 had no significant effects on fertility parameters (P > 0.05). However, animals immunized with Pep44 had lower egg fertilization rates (immunized 19.5% versus control 60.5%, P < 0.05) and produced significantly fewer embryos than control animals (immunized 0.5 embryos versus control 2.4 embryos, P < 0.05). The number of Pep44-immunized females that produced embryos was reduced by 64%. Identification and characterization of possum infertility-relevant epitopes on possum ZP2 protein will assist development of safe, humane, and possum-specific immunocontraceptive vaccines for controlling the introduced possums in New Zealand.

scholarly journals Molecular cloning and characterisation of alpha subunit of H+-ATPase in Lactobacillus casei Zhang

2009 ◽  
Vol 27 (No. 1) ◽  
pp. 49-54 ◽  
Author(s):  
X. Chen ◽  
M. Yang ◽  
Z. Sun ◽  
W. Liu ◽  
T. Sun ◽  
...  
Keyword(s):  
Amino Acids ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Inner Mongolia ◽  
Lactobacillus Casei ◽  
Cdna Sequence ◽  
Acid Tolerance ◽  
Alpha Subunit ◽  
Putative Protein ◽  
Atpase Gene

The acid tolerance is an important property of Lactic acid bacteria as potential probiotics. H<sup>+</sup>-ATPase is considered a key gene in several bacteria with the ability of acid tolerance. We cloned and sequenced the full length cDNA of alpha subunit of H<sup>+</sup>-ATPase gene in <i>Lactobacillus casei</i> Zhang, which had been isolated from traditional home-made koumiss in Inner Mongolia of China. The results showed that the respective cDNA sequence is composed of 1530 nucleotides and codes a putative protein including 509 amino acids. In addition, we also reconstructed the phylogenic trees for H<sup>+</sup>-ATPase gene based on amino acids sequences of diverse strains of Lactic acid bacteria.

scholarly journals Sequence and structure of mtr, an amino acid transport gene of Neurospora crassa

Genome ◽  
1991 ◽  
Vol 34 (4) ◽  
pp. 644-651 ◽  
Author(s):  
Kenneth Koo ◽  
W. Dorsey Stuart
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Neurospora Crassa ◽  
Rna Binding ◽  
Signal Sequence ◽  
Average Length ◽  
Open Reading Frame ◽  
Mrna Transcript ◽  
S1 Nuclease ◽  
Reading Frame

The gene product of the mtr locus of Neurospora crassa is required for the transport of neutral aliphatic and aromatic amino acids via the N system. We have previously cloned three cosmids containing Neurospora DNA that complement the mtr-6(r) mutant allele. The cloned DNAs were tightly linked to restriction fragment length polymorphisms that flank the mtr locus. A 2.9-kbp fragment from one cosmid was subcloned and found to complement the mtr-6(r) allele. Here we report the sequence of the fragment that hybridized to a poly(A)+ mRNA transcript of about 2300 nucleotides. We have identified an 845-bp open reading frame (ORF) having a 59-bp intron as the potential mtr ORF. S1 nuclease analysis of the transcript confirmed the transcript size and the presence of the intron. A second open reading frame was found upstream in the same reading frame as the mtr ORF and appears to be present in the mRNA transcript. The mtr ORF is predicted to encode a 261 amino acid polypeptide with a molecular mass of 28 613 Da. The proposed polypeptide exhibits six potential α-helical transmembrane domains with an average length of 23 amino acids, does not have a signal sequence, and contains amino acid sequence homologous to an RNA binding motif.Key words: sequence, membranes, ribonucleoprotein.

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
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