Cloning and characterization of an alternative splicing transcript of the gene coding for human cytidine deaminase

2007 ◽  
Vol 85 (1) ◽  
pp. 96-102 ◽  
Author(s):  
Bianca Cristina Garcia Lisboa ◽  
Tamara da Rocha Machado ◽  
Daniel Carvalho Pimenta ◽  
Sang Won Han
Keyword(s):  
Alternative Splicing ◽  
Dna Sequences ◽  
Genomic Sequence ◽  
Cytidine Deaminase ◽  
Alternative Form ◽  
Nih3t3 Cells ◽  
Reading Frame ◽  
Gene Coding ◽  
Identification And Characterization

Human cytidine deaminase (HCD) catalyzes the deamination of cytidine or deoxycytidine to uridine or deoxyuridine, respectively. The genomic sequence of HCD is formed by 31 kb with 4 exons and several alternative splicing signals, but an alternative form of HCD has yet to be reported. Here we describe the cloning and characterization of a small form of HCD, HSCD, and it is likely to be a product of alternative splicing of HCD. The alignment of DNA sequences shows that the HSCD matches HCD in 2 parts, except for a deletion of 170 bp. Based on the HCD genome organization, exons 1 and 4 should be joined and all sequences of introns and exons 2 and 3 should be deleted by splicing. This alternative splicing shifted the translation of the reading frame from the point of splicing. The estimated molecular mass is 9.8 kDa, and this value was confirmed by Western blot and mass spectroscopy after expressing the gene fused with glutathionine-S-transferase in the pGEX vector. The deletion and shift of the reading frame caused a loss of HCD activity, which was confirmed by enzyme assay and also with NIH3T3 cells modified to express HSCD and challenged against cytosine arabinoside. In this work we describe the identification and characterization of HSCD, which is the product of alternative splicing of the HCD gene.

scholarly journals Identification and characterization of a novel potyvirus infecting Paris yunnanensis

2021 ◽  
Author(s):  
Pingxiu Lan ◽  
Peng He ◽  
Mengji Cao ◽  
Guohua Zhou ◽  
Li Chenrong ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Genomic Sequence ◽  
Pairwise Comparison ◽  
Distinct Species ◽  
Reading Frame ◽  
Single Clade ◽  
Terminal Poly ◽  
Identification And Characterization ◽  
Complete Genomic

Abstract The complete genomic sequence of a novel potyvirus from Paris yunnanensis was determined by high-throughput sequencing then confirmed by Sanger sequencing. Its genomic RNA consists 9600 nucleotides (nt) excluding the 3’-terminal poly (A) tail, containing a typical large open reading frame (ORF) of potyviruses and encoding a putative polyprotein of 3098 amino acids (aa). Pairwise comparison analysis showed the virus shares sequence identity with other members of Potyvirus was 53.0–57.8% at genome sequence level, and 39.3–51.2% at polyprotein sequence level. Phylogenetic analysis indicated that the virus was clustered as a single clade within the genus Potyvirus both using nt and aa level. These results suggest that the virus should be considered as a distinct species within the genus Potyvirus, and it was tentatively named as “Paris mottle virus” (PaMoV).

scholarly journals Identification and Characterization of a Novel Potyvirus Infecting Paris Yunnanensis

2021 ◽  
Author(s):  
Pingxiu Lan ◽  
Peng He ◽  
Mengji Cao ◽  
Guohua Zhou ◽  
Chenrong Li ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Genomic Sequence ◽  
Pairwise Comparison ◽  
Distinct Species ◽  
Plum Pox Virus ◽  
Reading Frame ◽  
Terminal Poly ◽  
Chilli Veinal Mottle Virus ◽  
Identification And Characterization

Abstract The complete genomic sequence of a novel potyvirus from Paris yunnanensis was determined by high-throughput sequencing then confirmed by Sanger sequencing. Its genomic RNA consists 9600 nucleotides (nt) excluding the 3’-terminal poly (A) tail, containing a typical large open reading frame (ORF) of potyviruses and encoding a putative polyprotein of 3098 amino acids (aa). Pairwise comparison analysis showed the virus shares sequence identity with other members of Potyvirus was 53.0% to 57.8% at genome sequence level, and 39.3% to 51.2% at polyprotein sequence level. Phylogenetic analysis indicated that the virus was most closely related to the subgroup of plum pox virus and that of chilli veinal mottle virus within the genus Potyvirus. These results suggest that the virus should be considered as a distinct species within the genus Potyvirus and was tentatively named as “Paris mottle associated virus” (PMaV).

Characterization of Salt Overly Sensitive 1 (SOS1) gene homoeologs in quinoa (Chenopodium quinoa Willd.)

Genome ◽  
2009 ◽  
Vol 52 (7) ◽  
pp. 647-657 ◽  
Author(s):  
P. J. Maughan ◽  
T. B. Turner ◽  
C. E. Coleman ◽  
D. B. Elzinga ◽  
E. N. Jellen ◽  
...  
Keyword(s):  
Salt Tolerance ◽  
Dna Sequences ◽  
Genomic Sequence ◽  
Chenopodium Quinoa ◽  
Fish Analysis ◽  
Cdna Sequences ◽  
Grain Crop ◽  
High Level ◽  
Salt Overly Sensitive

Salt tolerance is an agronomically important trait that affects plant species around the globe. The Salt Overly Sensitive 1 (SOS1) gene encodes a plasma membrane Na+/H+ antiporter that plays an important role in germination and growth of plants in saline environments. Quinoa (Chenopodium quinoa Willd.) is a halophytic, allotetraploid grain crop of the family Amaranthaceae with impressive nutritional content and an increasing worldwide market. Many quinoa varieties have considerable salt tolerance, and research suggests quinoa may utilize novel mechanisms to confer salt tolerance. Here we report the cloning and characterization of two homoeologous SOS1 loci (cqSOS1A and cqSOS1B) from C. quinoa, including full-length cDNA sequences, genomic sequences, relative expression levels, fluorescent in situ hybridization (FISH) analysis, and a phylogenetic analysis of SOS1 genes from 13 plant taxa. The cqSOS1A and cqSOS1B genes each span 23 exons spread over 3477 bp and 3486 bp of coding sequence, respectively. These sequences share a high level of similarity with SOS1 homologs of other species and contain two conserved domains, a Nhap cation-antiporter domain and a cyclic-nucleotide binding domain. Genomic sequence analysis of two BAC clones (98 357 bp and 132 770 bp) containing the homoeologous SOS1 genes suggests possible conservation of synteny across the C. quinoa sub-genomes. This report represents the first molecular characterization of salt-tolerance genes in a halophytic species in the Amaranthaceae as well as the first comparative analysis of coding and non-coding DNA sequences of the two homoeologous genomes of C. quinoa.

scholarly journals Transcriptome-Based Identification of a Functional Fasciola hepatica Carboxylesterase B

Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1454
Author(s):  
Yaretzi J. Pedroza-Gómez ◽  
Raquel Cossio-Bayugar ◽  
Hugo Aguilar-Díaz ◽  
Silvana Scarcella ◽  
Enrique Reynaud ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Dna Sequences ◽  
Bioinformatics Analysis ◽  
Fasciola Hepatica ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cellular Membrane ◽  
Reading Frame ◽  
Protein Mass ◽  
Gene Coding ◽  
Sds Page

Bioinformatics analysis of the complete transcriptome of Fasciola hepatica, identified a total of ten putative carboxylesterase transcripts, including a 3146 bp mRNA transcript coding a 2205 bp open reading frame that translates into a protein of 735 amino acids, resulting in a predicted protein mass of 83.5 kDa and a putative carboxylesterase B enzyme. The gene coding for this enzyme was found in two reported F. hepatica complete genomes stretching 23,230 bp, containing two exons of 1282 and 1864 bp, respectively, as well as a 20,084 bp intron between the exons. The enzymatic activity was experimentally assayed on F. hepatica protein extracts by SDS-PAGE zymograms using synthetic chromogenic substrates, confirming both the theoretical molecular weight and carboxylesterase enzymatic activity. Further bioinformatics predicted that this enzyme is an integral component of the cellular membrane that should be active as a 167 kDa homodimer complex and polyacrylamide gel electrophoresis (PAGE) zymograms experiments confirmed the analysis. Additional bioinformatics analysis showed that DNA sequences that code for this particular enzyme are highly conserved in other parasitic trematodes, although they are labeled hypothetical proteins.

scholarly journals Survey, Identification, and Characterization of Cylindrocarpon-Like Asexual Morphs in Spanish Forest Nurseries

Plant Disease ◽  
2018 ◽  
Vol 102 (11) ◽  
pp. 2083-2100 ◽  
Author(s):  
Beatriz Mora-Sala ◽  
Ana Cabral ◽  
Maela León ◽  
Carlos Agustí-Brisach ◽  
Josep Armengol ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Elongation Factor ◽  
Its Region ◽  
Phylogenetic Position ◽  
Translation Elongation ◽  
Forest Nurseries ◽  
Asexual Morphs ◽  
Identification And Characterization ◽  
Elongation Factor 1

Cylindrocarpon-like asexual morphs infect herbaceous and woody plants, mainly in agricultural scenarios, but also in forestry systems. The aim of the present study was to characterize a collection of Cylindrocarpon-like isolates recovered from the roots of a broad range of forest hosts from nurseries showing decline by morphological and molecular studies. Between 2009 and 2012, 17 forest nurseries in Spain were surveyed and a total of 103 Cylindrocarpon-like isolates were obtained. Isolates were identified based on DNA sequences of the partial gene regions histone H3 (his3). For the new species, the internal transcribed spacer and intervening 5.8S nrRNA gene (ITS) region, β-tubulin (tub2), and translation elongation factor 1-α (tef1) were also used to determine their phylogenetic position. Twelve species belonging to the genera Cylindrodendrum, Dactylonectria, and Ilyonectria were identified from damaged roots of 15 different host genera. The species C. alicantinum, D. macrodidyma, D. novozelandica, D. pauciseptata, D. pinicola, D. torresensis, I. capensis, I. cyclaminicola, I. liriodendri, I. pseudodestructans, I. robusta, and I. rufa were identified. In addition, two Dactylonectria species (D. hispanica sp. nov. and D. valentina sp. nov.), one Ilyonectria species (I. ilicicola sp. nov.), and one Neonectria species (N. quercicola sp. nov.) are newly described. The present study demonstrates the prevalence of this fungal group associated with seedlings of diverse hosts showing decline symptoms in forest nurseries in Spain.

Identification and characterization of a gene coding a novel isoform of DEAD-box protein

2002 ◽  
Vol 14 (3) ◽  
pp. 185 ◽  
Author(s):  
Lanlan Yin ◽  
JianMin Li ◽  
Hu Zhu ◽  
Min Lin ◽  
Lijun Cheng ◽  
...  
Keyword(s):  
Human Testis ◽  
Microarray Hybridization ◽  
Chain Reaction ◽  
Dead Box ◽  
Gene Coding ◽  
Testis Cdna Library ◽  
Testis Cdna ◽  
Polymerase Chain ◽  
Identification And Characterization

A gene coding a novel isoform of DEAD-box protein named testicular DEAD-box protein (tDbp), presumably involved in testicular function, was identified and characterized. Testicular DEAD-box protein was cloned from a human testis cDNA library. The cDNA microarray hybridization showed that it was expressed at a higher level in adult testis than in embryo testis. Reverse transcription-polymerase chain reaction indicated that tDbp was specifically expressed in testis, but not in some other tissues.

Identification and characterization of G90, a novel mouse RNA that lacks an extensive open reading frame

Gene ◽  
1999 ◽  
Vol 232 (1) ◽  
pp. 35-42 ◽  
Author(s):  
Ralf Krause ◽  
Myriam Hemberger ◽  
Heinz Himmelbauer ◽  
Vera Kalscheuer ◽  
Reinald H. Fundele
Keyword(s):  
Open Reading Frame ◽  
Reading Frame ◽  
Identification And Characterization

scholarly journals Yos1p Is a Novel Subunit of the Yip1p–Yif1p Complex and Is Required for Transport between the Endoplasmic Reticulum and the Golgi Complex

2005 ◽  
Vol 16 (4) ◽  
pp. 1673-1683 ◽  
Author(s):  
Matthew Heidtman ◽  
Catherine Z. Chen ◽  
Ruth N. Collins ◽  
Charles Barlowe
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Secretory Pathway ◽  
Integral Membrane Protein ◽  
Rab Gtpases ◽  
Reading Frame ◽  
Multicopy Suppressor ◽  
Transport Vesicles ◽  
Identification And Characterization

Yeast Yip1p is a member of a conserved family of transmembrane proteins that interact with Rab GTPases. Previous studies also have indicated a role for Yip1p in the biogenesis of endoplasmic reticulum (ER)-derived COPII transport vesicles. In this report, we describe the identification and characterization of the uncharacterized open reading frame YER074W-A as a novel multicopy suppressor of the thermosensitive yip1-4 strain. We have termed this gene Yip One Suppressor 1 (YOS1). Yos1p is essential for growth and for function of the secretory pathway; depletion or inactivation of Yos1p blocks transport between the ER and the Golgi complex. YOS1 encodes an integral membrane protein of 87 amino acids that is conserved in eukaryotes. Yos1p localizes to ER and Golgi membranes and is efficiently packaged into ER-derived COPII transport vesicles. Yos1p associates with Yip1p and Yif1p, indicating Yos1p is a novel subunit of the Yip1p–Yif1p complex.

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