Isolation and Characterization of the First Putative Peroxidase Gene from Oilseed Rape (Brassica napus) which is Highly Homologous to HRPC

2006 ◽  
Vol 26 (3) ◽  
pp. 263-280 ◽  
Author(s):  
Weisheng Wu ◽  
Jie Lu ◽  
Yamin Wei ◽  
Jin Wang ◽  
Juan Lin ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Gene Family ◽  
Oilseed Rape ◽  
Blot Analysis ◽  
Reading Frame ◽  
Peroxidase Gene ◽  
Genomic Walking ◽  
Isolation And Characterization ◽  
New Gene ◽  
Rapid Amplification

A new gene, designated as BnPrx (GenBank Accession No. DQ078754), was isolated from oilseed rape (Brassica napus) by SMART Rapid Amplification of cDNA Ends (RACE). The full-length cDNA is 1307 bp long and contains a 1062 bp open reading frame (ORF), which encodes a 354 amino acid peroxidase precursor, with a 31 aa N-terminal signal peptide and a 15 aa C-terminal propeptide. The putative protein has a molecular weight of 38.86 kDa and a calculated pI of 5.85. BnPrx shares high identity with HRPC (89%). BnPrx possesses all active residues and two Ca2+ sites present in Horseradish peroxidase isoenzymes C (HRPC) as well as six N-glycosylation sites. The predicted 3-D structure of BnPrx is very similar to that of HRPC. Assisted by genomic walking technology, the genomic DNA of BnPrx was also cloned, consisting of 3 introns and 4 exons. Thirty-two TATA boxes, 18 CAAT boxes and many cis-elements, such as WUN, MeJR, were found in its promoter region. Southern blot analysis indicated that BnPrx belonged to a small gene family. Northern blot analysis revealed that BnPrx was constitutively expressed in all tested tissues, including roots, stems and leaves, with the high expression in leaves and stems. The expression of BnPrx could be induced by methyl jasmonate (MeJA), salicylic acid (SA), cold and H2O2. The cloning and characterizing of BnPrx might not only help us understand the physiological function and molecular evolution of the large peroxidase gene family more comprehensively, but also provide an alternative way of seeking a more effective and economical substitute for HRPC.

The extensin gene family in oilseed rape (Brassica napus L.): Characterisation of sequences of representative members of the family

1990 ◽  
Vol 223 (2) ◽  
pp. 273-287 ◽  
Author(s):  
I. Marta Evans ◽  
Laurence N. Gatehouse ◽  
John A. Gatehouse ◽  
Jennifer N. Yarwood ◽  
Donald Boulter ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Gene Family ◽  
Oilseed Rape ◽  
Brassica Napus L ◽  
The Family

Genome-Wide Identification and Evolutionary Analysis of the Fruit-Weight 2.2-Like Gene Family in Polyploid Oilseed Rape (Brassica napus L.)

2020 ◽  
Vol 39 (5) ◽  
pp. 766-782
Author(s):  
Chen Kuang ◽  
Jun Li ◽  
Hongfang Liu ◽  
Jun Liu ◽  
Xingchao Sun ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Gene Family ◽  
Oilseed Rape ◽  
Fruit Weight ◽  
Evolutionary Analysis ◽  
Brassica Napus L ◽  
Genome Wide

scholarly journals Isolation and characterization of SUA5, a novel gene required for normal growth in Saccharomyces cerevisiae.

Genetics ◽  
1992 ◽  
Vol 131 (4) ◽  
pp. 791-801 ◽  
Author(s):  
J G Na ◽  
I Pinto ◽  
M Hampsey
Keyword(s):  
Transcription Initiation ◽  
State Level ◽  
Normal Growth ◽  
Gene Encoding ◽  
Reading Frame ◽  
Isolation And Characterization ◽  
New Gene ◽  
Type Gene ◽  
Data Banks

Abstract We have identified the sua5 locus as a suppressor of an aberrant ATG codon located in the leader region of the cyc1 gene. The sua5-1 allele enhances the iso-1-cytochrome c steady state level in the cyc1-1019 mutant from 2% to approximately 60% of normal (Cyc+) and also confers a marked slow growth (Slg-) phenotype. Suppression is not a consequence of altered transcription initiation at the cyc1 locus. The SUA5 wild-type gene was isolated and sequenced, revealing an open reading frame (ORF) encoding a potential protein of 46,537 Da. SUA5 transcript analyses were consistent with expression of the predicted ORF and Sua5 antisera detected a protein with an apparent molecular mass of 44 kDa. SUA5 was mapped to chromosome VII, immediately adjacent to the PMR1 gene. Hybridization analysis revealed the presence of a related gene on chromosome XII. Neither the SUA5 DNA sequence nor deduced amino acid sequence showed homology to any sequences in the data banks. Disruption of SUA5 conferred the same Cyc+ and Slg- phenotypes as the sua5-1 suppressor, which is the result of a missense mutation, encoding a Ser107----Phe replacement. In addition, sua5 null mutants lack cytochrome a.a3 and fail to grow on lactate or glycerol medium. These results define SUA5 as a new gene encoding a novel protein that is necessary for normal cell growth.

scholarly journals Isolation and Characterization of the Betaine Aldehyde Dehydrogenase Gene in

2010 ◽  
Vol 4 (1) ◽  
pp. 18-25 ◽  
Author(s):  
Jun Liu ◽  
Huiming Zeng ◽  
Xue Li ◽  
Lixin Xu ◽  
Yingbo Wang ◽  
...  
Keyword(s):  
Aldehyde Dehydrogenase ◽  
Glycine Betaine ◽  
Betaine Aldehyde Dehydrogenase ◽  
Reading Frame ◽  
Betaine Aldehyde ◽  
Isolation And Characterization ◽  
Dehydrogenase Gene ◽  
Ophiopogon Japonicus ◽  
Rapid Amplification

Betaine aldehyde dehydrogenase (BADH) catalyzes the last step in the synthesis of the glycine betaine from choline. The BADH gene from turfgrass Ophiopogon japonicus has not been reported. In this study, we first isolated the full length cDNA of betaine aldehyde dehydrogenase gene (OjBADH) from O. japonicus using Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR) and Rapid Amplification of cDNA Ends (RACE) techniques. The OjBADH gene (GenBank accession number: DQ645888) has 1785 nucleotides with the 5’ untranscribed region (UTR) of 63 nucleotides, 3’ UTR of 219 nucleotides, and an open reading frame of 1503 nucleotides. This gene encodes a polypeptide of 500 amino acids. It shares a high homology with BADH genes of other Chenopodiaceae species. The putative protein includes a conservative region of phosphofructokinase, aldehyde dehydrogenase, and glutamy phosphoric acid reductase. Overexpression of OjBADH in transgenic tobacco plants demonstrated 2-2.5 folds increase of glycine betaine content and 60- 85% increase of survival rate under salt tolerance. These results suggested that the O. japonicus BADH gene may be used to engineer plants for salt stress tolerance.

Isolation and characterization of an oilseed rape SKP1 gene BnSKP1 involved on defense in Brassica napus

2008 ◽  
Vol 136 ◽  
pp. S227 ◽  
Author(s):  
Heng Yin ◽  
Shuguang Li ◽  
Xiaoming Zhao ◽  
XueFang Bai ◽  
YuGuang Du
Keyword(s):  
Brassica Napus ◽  
Oilseed Rape ◽  
Isolation And Characterization

Molecular cloning and expression of a cDNA encoding the proliferating cell nuclear antigen from Brassica napus (oilseed rape)

Genome ◽  
1993 ◽  
Vol 36 (3) ◽  
pp. 459-466 ◽  
Author(s):  
Nancy-Ann Markley ◽  
Peta C. Bonham-Smith ◽  
Maurice M. Moloney
Keyword(s):  
Dna Replication ◽  
Brassica Napus ◽  
Plant Cell ◽  
Oilseed Rape ◽  
Proliferating Cell Nuclear Antigen ◽  
Higher Plants ◽  
Cloning And Expression ◽  
Nuclear Antigen ◽  
Reading Frame ◽  
Proliferating Cell

A cDNA clone encoding the proliferating cell nuclear antigen (PCNA) has been isolated from a Brassica napus apical meristem cDNA library. The putative full-length cDNA contains an open reading frame of 1004 nucleotides, which predicts a protein of 263 amino acids (Mr = 29 231). Sequence analysis has revealed that the plant PCNA exhibits 81.6% amino acid similarity with the human PCNA. Genomic Southern blot analysis indicates the presence of at least two copies of PCNA per genome. The B. napus PCNA mRNA (1.0 kb) was expressed in rapidly dividing tissues such as flower buds, apical meristems, and young leaves, while mature stem and fully expanded leaves showed significantly lower levels of PCNA transcript. The B. napus PCNA cDNA was expressed in Escherichia coli as a fusion protein with glutathione-5-transferase (GST) in the bacterial expression vector pGEX-2T. A broad specificity monoclonal antibody raised against rabbit PCNA cross-reacted with the GST–PCNA fusion peptide but not with the GST moiety alone. This antibody also recognized the human PCNA (36 kDa) polypeptide, confirming the structural similarities between the human and plant PCNA. The high degree of structural conservation of PCNA from such diverse organisms as humans and higher plants suggests that the plant PCNA may function in a manner analogous to that found in mammals with respect to plant cell DNA replication. Such conservation suggests that PCNA is also a critical component of the plant cell DNA replication complex.Key words: proliferating cell nuclear antigen, DNA replication, DNA polymerase-δ, cell cycle regulation, Brassica napus (oilseed rape).

Genome-wide analysis and expression profiling of the GRF gene family in oilseed rape ( Brassica napus L.)

Gene ◽  
2017 ◽  
Vol 620 ◽  
pp. 36-45 ◽  
Author(s):  
Jin-Qi Ma ◽  
Hong-Ju Jian ◽  
Bo Yang ◽  
Kun Lu ◽  
Ao-Xiang Zhang ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Gene Family ◽  
Oilseed Rape ◽  
Expression Profiling ◽  
Brassica Napus L ◽  
Genome Wide Analysis ◽  
Genome Wide

scholarly journals Functional and molecular identification of a TASK-1 potassium channel regulating chloride secretion through CFTR channels in the shark rectal gland: implications for cystic fibrosis

2016 ◽  
Vol 311 (6) ◽  
pp. C884-C894 ◽  
Author(s):  
Connor J. Telles ◽  
Sarah E. Decker ◽  
William W. Motley ◽  
Alexander W. Peters ◽  
Ali Poyan Mehr ◽  
...  
Keyword(s):  
Potassium Channel ◽  
Reversal Potential ◽  
Chloride Secretion ◽  
Rectal Gland ◽  
Degenerate Primers ◽  
Reading Frame ◽  
Genomic Walking ◽  
Shark Rectal Gland ◽  
Mammalian Lung ◽  
Rapid Amplification

In the shark rectal gland (SRG), apical chloride secretion through CFTR channels is electrically coupled to a basolateral K+ conductance whose type and molecular identity are unknown. We performed studies in the perfused SRG with 17 K+ channel inhibitors to begin this search. Maximal chloride secretion was markedly inhibited by low-perfusate pH, bupivicaine, anandamide, zinc, quinidine, and quinine, consistent with the properties of an acid-sensitive, four-transmembrane, two-pore-domain K+ channel (4TM-K2P). Using PCR with degenerate primers to this family, we identified a TASK-1 fragment in shark rectal gland, brain, gill, and kidney. Using 5′ and 3′ rapid amplification of cDNA ends PCR and genomic walking, we cloned the full-length shark gene (1,282 bp), whose open reading frame encodes a protein of 375 amino acids that was 80% identical to the human TASK-1 protein. We expressed shark and human TASK-1 cRNA in Xenopus oocytes and characterized these channels using two-electrode voltage clamping. Both channels had identical current-voltage relationships (outward rectifying) and a reversal potential of −90 mV. Both were inhibited by quinine, bupivicaine, and acidic pH. The pKa for current inhibition was 7.75 for shark TASK-1 vs. 7.37 for human TASK-1, values similar to the arterial pH for each species. We identified this protein in SRG by Western blot and confocal immunofluorescent microscopy and detected the protein in SRG and human airway cells. Shark TASK-1 is the major K+ channel coupled to chloride secretion in the SRG, is the oldest 4TM 2P family member identified, and is the first TASK-1 channel identified to play a role in setting the driving force for chloride secretion in epithelia. The detection of this potassium channel in mammalian lung tissue has implications for human biology and disease.

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