scholarly journals CMYB1Encoding a MYB Transcriptional Activator Is Involved in Abiotic Stress and Circadian Rhythm in Rice

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Min Duan ◽  
Peng Huang ◽  
Xi Yuan ◽  
Hui Chen ◽  
Ji Huang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcriptional Activation ◽  
Fluorescent Protein ◽  
Developmental Stages ◽  
Transcriptional Activator ◽  
Myb Transcription Factor ◽  
Specific Gene ◽  
Protein Fusion ◽  
Rhythmic Expression ◽  
Green Fluorescent

Through analysis of cold-induced transcriptome, a novel gene encoding a putative MYB transcription factor was isolated and designatedCold induced MYB 1 (CMYB1). Tissue-specific gene expression analysis revealed thatCMYB1was highly expressed in rice stems and nodes. qRT-PCR assay indicated thatCMYB1was dramatically induced by cold stress (>100-folds) and induced by exogenous ABA and osmotic stress. Interestingly,CMYB1showed rhythmic expression profile in rice leaves at different developmental stages. Subcellular localization assay suggested thatCMYB1-GFP (green fluorescent protein) fusion protein was localized in the nuclei. Moreover,CMYB1exhibited the transcriptional activation activity when transiently expressed in rice protoplast cells. Taken together,CMYB1probably functions as a transcriptional activator in mediating stress and rhythm responsive gene expression in rice.

In Situ Gene Expression in Mixed-Culture Biofilms: Evidence of Metabolic Interactions between Community Members

1998 ◽  
Vol 64 (2) ◽  
pp. 721-732 ◽  
Author(s):  
Søren Møller ◽  
Claus Sternberg ◽  
Jens Bo Andersen ◽  
Bjarke Bak Christensen ◽  
Juan Luis Ramos ◽  
...  
Keyword(s):  
Gene Expression ◽  
Microbial Communities ◽  
Pure Culture ◽  
Fluorescent Protein ◽  
Confocal Laser ◽  
Specific Gene ◽  
Community Members ◽  
Metabolic Interactions ◽  
Green Fluorescent

ABSTRACT Microbial communities growing in laboratory-based flow chambers were investigated in order to study compartmentalization of specific gene expression. Among the community members studied, the focus was in particular on Pseudomonas putida and a strain of anAcinetobacter sp., and the genes studied are involved in the biodegradation of toluene and related aromatic compounds. The upper-pathway promoter (Pu) and themeta-pathway promoter (Pm) from the TOL plasmid were fused independently to the gene coding for the green fluorescent protein (GFP), and expression from these promoters was studied inP. putida, which was a dominant community member. Biofilms were cultured in flow chambers, which in combination with scanning confocal laser microscopy allowed direct monitoring of promoter activity with single-cell spatial resolution. Expression from thePu promoter was homogeneously induced by benzyl alcohol in both community and pure-culture biofilms, while the Pmpromoter was induced in the mixed community but not in a pure-culture biofilm. By sequentially adding community members, induction ofPm was shown to be a consequence of direct metabolic interactions between an Acinetobacter species and P. putida. Furthermore, in fixed biofilm samples organism identity was determined and gene expression was visualized at the same time by combining GFP expression with in situ hybridization with fluorescence-labeled 16S rRNA targeting probes. This combination of techniques is a powerful approach for investigating structure-function relationships in microbial communities.

scholarly journals Nanoluciferase-Based Method for Detecting Gene Expression in Caenorhabditis elegans

Genetics ◽  
2019 ◽  
Vol 213 (4) ◽  
pp. 1197-1207 ◽  
Author(s):  
Ivana Sfarcic ◽  
Theresa Bui ◽  
Erin C. Daniels ◽  
Emily R. Troemel
Keyword(s):  
Gene Expression ◽  
Caenorhabditis Elegans ◽  
Promoter Activity ◽  
Fluorescent Protein ◽  
Developmental Stages ◽  
Fluorescent Reporters ◽  
C Elegans ◽  
Link Type ◽  
Highly Sensitive ◽  
Green Fluorescent

Genetic reporters such as the green fluorescent protein (GFP) can facilitate measurement of promoter activity and gene expression. However, animal autofluorescence limits the sensitivity of GFP and other fluorescent reporters in whole-animal settings like in the nematode Caenorhabditis elegans. Here, we present a highly sensitive Nanoluciferase (NanoLuc)-based method in a multiwell format to detect constitutive and inducible gene expression in C. elegans. We optimize detection of bioluminescent signals from NanoLuc in C. elegans and show that it can be detected at 400,000-fold over background in a population of 100 animals expressing intestinal NanoLuc driven by the vha-6 promoter. We can reliably detect signal in single vha-6p::Nanoluc-expressing worms from all developmental stages. Furthermore, we can detect signal from a 1/100 dilution of lysate from a single vha-6p::Nanoluc-expressing adult and from a single vha-6p::Nanoluc-expressing adult “hidden” in a pool of 5000 N2 wild-type animals. We also optimize various steps of this protocol, which involves a lysis step that can be performed in minutes. As a proof-of-concept, we used NanoLuc to monitor the promoter activity of the pals-5 stress/immune reporter and were able to measure 300- and 50-fold increased NanoLuc activity after proteasome blockade and infection with microsporidia, respectively. Altogether, these results indicate that NanoLuc provides a highly sensitive genetic reporter for rapidly monitoring whole-animal gene expression in C. elegans.

scholarly journals Nanoluciferase-based Method for Detecting Gene Expression inC. elegans

2019 ◽  
Author(s):  
Ivana Sfarcic ◽  
Theresa Bui ◽  
Erin C. Daniels ◽  
Emily R. Troemel
Keyword(s):  
Gene Expression ◽  
Promoter Activity ◽  
Fluorescent Protein ◽  
Developmental Stages ◽  
C Elegans ◽  
Highly Sensitive ◽  
Monitoring Gene Expression ◽  
Green Fluorescent ◽  
Limited Sensitivity ◽  
Inducible Gene

AbstractGenetic reporters such as the green fluorescent protein (GFP) can facilitate measurement of promoter activity and gene expression. However, GFP and other fluorophores have limited sensitivity, particularly in tissues that autofluoresce like the intestine of the nematodeCaenorhabditis elegans. Here, we present a highly sensitive Nanoluciferase (NanoLuc)-based method in multi-well format to detect constitutive and inducible gene expression inC. elegans. We optimize detection of bioluminescent signal from NanoLuc inC. elegansand show that it can be detected at 400,000-fold over background in a population of 100 animals expressing intestinal NanoLuc driven by thevha-6promoter. We can reliably detect signal in singlevha-6p::Nanoluc-expressing worms from all developmental stages. Furthermore, we can detect signal from 1/100 dilution of lysate from a singlevha-6p::Nanoluc-expressing adult and from a singlevha-6p::Nanoluc-expressing adult “hidden” in a pool of 5,000 N2 wild-type animals. We also optimized various steps of this protocol, which involves a lysis step that can be performed in minutes. As a proof of concept, we used NanoLuc to monitor promoter activity of thepals-5stress/immune reporter and we were able to measure 300 and 50-fold increased NanoLuc activity after proteasome blockade and infection with microsporidia, respectively. Altogether, these results indicate that NanoLuc provides a highly sensitive genetic reporter for rapidly monitoring gene expression inC. elegans.

scholarly journals Role of an FtsK-Like Protein in Genetic Stability in Streptomyces coelicolor A3(2)

2007 ◽  
Vol 189 (6) ◽  
pp. 2310-2318 ◽  
Author(s):  
Lei Wang ◽  
Yanfei Yu ◽  
Xinyi He ◽  
Xiufen Zhou ◽  
Zixin Deng ◽  
...  
Keyword(s):  
Cell Division ◽  
Chromosome Segregation ◽  
Fluorescent Protein ◽  
Streptomyces Coelicolor ◽  
Complex Life Cycle ◽  
Specific Gene ◽  
Chromosomal Dna ◽  
Protein Fusion ◽  
Fluorescent Reporter ◽  
Green Fluorescent

ABSTRACT Streptomyces coelicolor A3(2) does not have a canonical cell division cycle during most of its complex life cycle, yet it contains a gene (ftsKSC ) encoding a protein similar to FtsK, which couples the completion of cell division and chromosome segregation in unicellular bacteria such as Escherichia coli. Here, we show that various constructed ftsKSC mutants all grew apparently normally and sporulated but upon restreaking gave rise to many aberrant colonies and to high frequencies of chloramphenicol-sensitive mutants, a phenotype previously associated with large terminal deletions from the linear chromosome. Indeed, most of the aberrant colonies had lost large fragments near one or both chromosomal termini, as if chromosome ends had failed to reach their prespore destination before the closure of sporulation septa. A constructed FtsKSC-enhanced green fluorescent protein fusion protein was particularly abundant in aerial hyphae, forming distinctive complexes before localizing to each sporulation septum, suggesting a role for FtsKSC in chromosome segregation during sporulation. Use of a fluorescent reporter showed that when ftsKSC was deleted, several spore compartments in most spore chains failed to express the late-sporulation-specific sigma factor gene sigF, even though they contained chromosomal DNA. This suggested that sigF expression is autonomously activated in each spore compartment in response to completion of chromosome transfer, which would be a previously unknown checkpoint for late-sporulation-specific gene expression. These results provide new insight into the genetic instability prevalent among streptomycetes, including those used in the industrial production of antibiotics.

Sign in / Sign up

Export Citation Format

Share Document