Characterization of the mitochondrial orfB gene and its derivative, orf224, a chimeric open reading frame specific to one mitochondrial genome of the ?Polima? male-sterile cytoplasm in rapeseed (Brassica napus L.)

1995 ◽  
Vol 28 (6) ◽  
pp. 546-552 ◽  
Author(s):  
Hirokazu Handa ◽  
Jos� Manuel Gualberto ◽  
Jean-Michel Grienenberger
Keyword(s):  
Brassica Napus ◽  
Mitochondrial Genome ◽  
Open Reading Frame ◽  
Male Sterile ◽  
Brassica Napus L ◽  
Reading Frame ◽  
Male Sterile Cytoplasm

scholarly journals Characterization of the BnA10.tfl1 Gene Controls Determinate Inflorescence Trait in Brassica napus L.

Agronomy ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 722 ◽  
Author(s):  
Yongpeng Jia ◽  
Kaixiang Li ◽  
Haidong Liu ◽  
Lingxiong Zan ◽  
Dezhi Du
Keyword(s):  
Brassica Napus ◽  
Shoot Apex ◽  
Apical Meristem ◽  
Brassica Napus L ◽  
Reading Frame ◽  
Terminal Flower ◽  
Terminal Flower 1 ◽  
Oilseed Breeding ◽  
Phosphatidylethanolamine Binding Protein

Determinate inflorescences have a significant effect on the genetic improvement of rapeseed, so understanding the molecular function underlying the inflorescence trait may be beneficial to oilseed breeding. A previous study found candidate gene BnTFL1 (Terminal Flower 1) for control of the inflorescence trait on Brassica napus chromosome A10 (16,627–16,847 kb). However, little is known about the function of the BnTFL1 gene in B. napus. In this study, we firstly studied the formation of the shoot apical meristem and gene expression in indeterminate and determinate inflorescences; the results showed that the inflorescence architecture and BnA10.TFL1 expression showed significant differences in the shoot apex at the budding stage. Then, two alleles (named BnA10.TFL1 gene from indeterminate and BnA10.tfl1 gene from determinate) were cloned and sequence-analyzed; the results suggest that the open reading frame of the alleles comprises 537 bp, encodes 178 amino acids containing a conserved phosphatidylethanolamine-binding protein (PEBP) domain, and shares high similarity with Arabidopsis thaliana TFL1. To analyze the function of BnA10.TFL1, the BnA10.TFL1 gene was introduced into the determinate A. thaliana tfl1 mutant and B. napus 571 line by complementation experiment. The determinate traits were restored to indeterminate, and expression of BnA10.TFL1 was increased in the indeterminate shoot apex. These results reveal that BnA10.tfl1 is a gene controlling the determinate inflorescence trait. Moreover, the BnA10.TFL1 protein was localized to the nucleus, cytoplasm, and plasma membrane. Collectively, the results of this study help us to understand the molecular mechanism of determinate inflorescences and will provide a reliable research basis for the application of determinate inflorescences in B. napus.

Characterization of a new temperature-sensitive male sterile line SP2S in rapeseed (Brassica napus L.)

Euphytica ◽  
2015 ◽  
Vol 206 (2) ◽  
pp. 473-485 ◽  
Author(s):  
Chengyu Yu ◽  
Yingfen Guo ◽  
Juan Ge ◽  
Yumei Hu ◽  
Jungang Dong ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Temperature Sensitive ◽  
Male Sterile ◽  
Male Sterile Line ◽  
Brassica Napus L

Comparison of the effect of nap and pol cytoplasms on the performance of three summer oilseed rape cultivar-derived isoline pairs

1994 ◽  
Vol 74 (4) ◽  
pp. 729-731 ◽  
Author(s):  
P. B. E. McVetty ◽  
R. Pinnisch
Keyword(s):  
Brassica Napus ◽  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Oil Content ◽  
Relative Effect ◽  
Male Sterile ◽  
Negative Effects ◽  
Brassica Napus L ◽  
Male Sterile Cytoplasm ◽  
The One

The pol cytoplasm is a male sterile cytoplasm with potential for use in hybrid summer rape (Brassica napus L.) seed production while the nap cytoplasm is the one most commonly encountered in summer rape cultivars. The objective of this study was to compare the performance of three cultivar-derived summer rape isoline pairs in the nap and pol cytoplasms to determine the relative effect on performance of these two cytoplasms. One nap line yielded significantly more than its corresponding pol line, three nap lines had significantly higher oil content than their corresponding pol lines, two nap lines had significantly higher protein content than their corresponding pol lines, and two nap lines produced significantly more seed energy than their corresponding pol lines. There are pleiotropic negative effects (biological costs) associated with the pol cytoplasm. These negative effects are affected by nuclear genotype and appear to be related to the depth of male sterility expressed in the derived pol A-line. Key words: Cytoplasm cost, Brassica napus L., cytoplasmic male sterility

scholarly journals Antigenic Characterization of ORF2 and ORF3 Proteins of Hepatitis E Virus (HEV)

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1385
Author(s):  
Giulia Pezzoni ◽  
Lidia Stercoli ◽  
Eleonora Pegoiani ◽  
Emiliana Brocchi
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Recombinant Antigen ◽  
Open Reading Frame ◽  
Reading Frame ◽  
E Coli ◽  
Antigenic Characterization ◽  
Antigenic Properties ◽  
Open Reading Frame 2

To evaluate the antigenic properties of Hepatitis E Virus (HEV) Open Reading Frame 2 and 3 (ORF2 and ORF3) codified proteins, we expressed different portions of ORF2 and the entire ORF3 in E. coli, a truncated ORF2, was also expressed in baculovirus. A panel of 37 monoclonal antibodies (MAbs) was raised against ORF2 (1–660 amino acids) and MAbs were mapped and characterized using the ORF2 expressed portions. Selected HEV positive and negative swine sera were used to evaluate ORF2 and ORF3 antigens’ immunogenicity. The MAbs were clustered in six groups identifying six antigenic regions along the ORF2. Only MAbs binding to the sixth ORF2 antigenic region (394–608 aa) were found to compete with HEV positive sera and efficiently catch the recombinant antigen expressed in baculovirus. The ORF2 portion from 394–608 aa demonstrated to include most immunogenic epitopes with 85% of HEV positive swine sera reacting against the region from 461–544 aa. Only 5% of the selected HEV sera reacted against the ORF3 antigen.

scholarly journals Gene Cloning and Molecular Characterization of a Two-Enzyme System Catalyzing the Oxidative Detoxification of β-Endosulfan

2002 ◽  
Vol 68 (12) ◽  
pp. 6237-6245 ◽  
Author(s):  
Tara D. Sutherland ◽  
Irene Horne ◽  
Robyn J. Russell ◽  
John G. Oakeshott
Keyword(s):  
Shuttle Vector ◽  
Open Reading Frame ◽  
Cosmid Library ◽  
Flavin Mononucleotide ◽  
Flavin Reductase ◽  
Reading Frame ◽  
Reductase Gene ◽  
Degradation Activity ◽  
Layer Chromatography

ABSTRACT The gram-positive bacterium Mycobacterium sp. strain ESD is able to use the cyclodiene insecticide endosulfan as a source of sulfur for growth. This activity is dependent on the absence of sulfite or sulfate in the growth medium. A cosmid library of strain ESD DNA was constructed in a Mycobacterium-Escherichia coli shuttle vector and screened for endosulfan-degrading activity in Mycobacterium smegmatis, a species that does not degrade endosulfan. Using this method, we identified a single cosmid that conferred sulfur-dependent endosulfan-degrading activity on the host strain. An open reading frame (esd) was identified within this cosmid that, when expressed behind a constitutive promoter in a mycobacterial expression vector, conferred sulfite- and sulfate-independent β-endosulfan degradation activity on the recombinant strain. The translation product of this gene (Esd) had up to 50% sequence identity with an unusual family of monooxygenase enzymes that use reduced flavins, provided by a separate flavin reductase enzyme, as cosubstrates. An additional partial open reading frame was located upstream of the Esd gene that had sequence homology to the same monooxygenase family. A flavin reductase gene, identified in the M. smegmatis genome, was cloned, expressed, and used to provide reduced flavin mononucleotide for Esd in enzyme assays. Thin-layer chromatography and gas chromatography analyses of the enzyme assay mixtures revealed the disappearance of β-endosulfan and the appearance of the endosulfan metabolites, endosulfan monoaldehyde and endosulfan hydroxyether. This suggests that Esd catalyzes the oxygenation of β-endosulfan to endosulfan monoaldehyde and endosulfan hydroxyether. Esd did not degrade either α-endosulfan or the metabolite of endosulfan, endosulfan sulfate.

scholarly journals Cloning and characterization of α-pinene synthase from Chamaecyparis formosensis Matsum

Holzforschung ◽  
2009 ◽  
Vol 63 (1) ◽  
Author(s):  
Fang-Hua Chu ◽  
Pei-Min Kuo ◽  
Yu-Rong Chen ◽  
Sheng-Yang Wang
Keyword(s):  
Major Product ◽  
Open Reading Frame ◽  
Terpene Synthase ◽  
Geranyl Diphosphate ◽  
Reading Frame ◽  
E Coli ◽  
Phylogenetic Taxonomy ◽  
Chamaecyparis Formosensis ◽  
Pcr Method

AbstractAnalyzing the gene sequences of terpene synthase (TPS) may contribute to a better understanding of terpenes biosynthesis and evolution of phylogenetic taxonomy.Chamaecyparis formosensisis an endemic and precious conifer of Taiwan. To understand the biosynthesis mechanism of terpenes in this tree, a full length of putative mono-TPS, named asCf-Pin(GeneBank accession no. EU099434), was obtained by PCR method and RACE extension. TheCf-Pinhas an 1887-bp open reading frame and encodes 628 amino acids. To identify the function ofCf-Pin,the recombinant protein fromEscherichia coliwas incubated with geranyl diphosphate, produced one major product, the structure of which was elucidated. GC/MS analysis and matching of retention time and mass spectrum with authentic standards revealed that this product isα-pinene. This is the first report of cloning of a mono-TPS and functionally expressed inE. coliand which could be identified asα-pinene synthase from a Cupressaceae conifer.

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