Amylase gene expression in intraspecific and interspecific somatic transformants of Drosophila

Genome ◽  
1990 ◽  
Vol 33 (4) ◽  
pp. 501-508 ◽  
Author(s):  
Sylvia A. Hawley ◽  
R. A. Norman ◽  
Celeste J. Brown ◽  
Winifred W. Doane ◽  
Wyatt W. Anderson ◽  
...  
Keyword(s):  
Functional Status ◽  
Transient Expression ◽  
Internal Control ◽  
Relative Activity ◽  
Activity Levels ◽  
Transient Expression Assay ◽  
Homologous Sequences ◽  
Electrophoretic Mobilities ◽  
Somatic Transformation ◽  
Amy Genes

The Amylase locus in Drosophila melanogaster normally contains two copies of the structural gene for α-amylase, a centromere-proximal copy, Amy-p, and a distal copy, Amy-d. Products of the two genes may display discrete electrophoretic mobilities, but many strains known to carry the Amy duplication are characterized by a single amylase electromorph, e.g., Oregon-R, which produces the mobility variant AMY-1. A transient expression assay was used in somatic transformation experiments to test the functional status of the Amy genes from an Oregon-R strain. Plasmid constructs containing either the proximal or distal copy were tested in amylase-null hosts. Both genes produced a functional AMY-1 isozyme. Constructs were tested against an AMY-3 reference activity produced by a coinjected plasmid that contains the Amy-d3 allele from a Canton-S strain. With reference to the internal control, the Amy-p and Amy-d genes from Oregon-R expressed different relative activity levels for AMY-1 in transient assays. The transient expression assay was successfully used to test the functional status of, Amy-homologous sequences from strains of other species of Drosophila characterized by a single amylase electromorph, namely, Drosophila pseudoobscura ST and Drosophila miranda S 204. The amylase-null strain of D. melanogaster provided the hosts for these interspecific somatic transformation experiments.Key words: α-amylase, Amy, transient assay, gene duplication, intergenic transformation.

A mechanism by which adenovirus virus-associated RNAI controls translation in a transient expression assay

1987 ◽  
Vol 7 (1) ◽  
pp. 549-551
Author(s):  
G Akusjärvi ◽  
C Svensson ◽  
O Nygård
Keyword(s):  
Transient Expression ◽  
Polypeptide Chain ◽  
Adenovirus Type ◽  
Initiation Factor ◽  
Translational Efficiency ◽  
Dna Transfection ◽  
Transient Expression Assay ◽  
293 Cells ◽  
Initiation Factor 2

The mechanism by which adenovirus virus-associated RNAI stimulates translational efficiency in a transient-expression assay in 293 cells was investigated. We showed that DNA transfection leads to activation of a protein kinase that phosphorylates the alpha subunit of eucaryotic initiation factor 2 and, as a consequence, inhibition of polypeptide chain initiation. Cotransfection of a plasmid encoding adenovirus type 2 virus-associated RNAI recovered the translational capacity by preventing activation of the kinase.

The phylogenetically invariant ACAGAGA and AGC sequences of U6 small nuclear RNA are more tolerant of mutation in human cells than in Saccharomyces cerevisiae

1993 ◽  
Vol 13 (9) ◽  
pp. 5377-5382
Author(s):  
B Datta ◽  
A M Weiner
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Transient Expression ◽  
Human Cells ◽  
Mrna Splicing ◽  
Small Nuclear Rna ◽  
Transient Expression Assay ◽  
U6 Snrna ◽  
Nuclear Rna

U6 small nuclear RNA (snRNA) is the most highly conserved of the five spliceosomal snRNAs that participate in nuclear mRNA splicing. The proposal that U6 snRNA plays a key catalytic role in splicing [D. Brow and C. Guthrie, Nature (London) 337:14-15, 1989] is supported by the phylogenetic conservation of U6, the sensitivity of U6 to mutation, cross-linking of U6 to the vicinity of the 5' splice site, and genetic evidence for extensive base pairing between U2 and U6 snRNAs. We chose to mutate the phylogenetically invariant 41-ACAGAGA-47 and 53-AGC-55 sequences of human U6 because certain point mutations within the homologous regions of Saccharomyces cerevisiae U6 selectively block the first or second step of mRNA splicing. We found that both sequences are more tolerant to mutation in human cells (assayed by transient expression in vivo) than in S. cerevisiae (assayed by effects on growth or in vitro splicing). These differences may reflect different rate-limiting steps in the particular assays used or differential reliance on redundant RNA-RNA or RNA-protein interactions. The ability of mutations in U6 nucleotides A-45 and A-53 to selectively block step 2 of splicing in S. cerevisiae had previously been construed as evidence that these residues might participate directly in the second chemical step of splicing; an indirect, structural role seems more likely because the equivalent mutations have no obvious phenotype in the human transient expression assay.

scholarly journals Functional status measures for the COPD patient: A practical categorization

2018 ◽  
Vol 16 ◽  
pp. 147997311881646 ◽  
Author(s):  
Suzanne Claire Lareau ◽  
Felicity Clair Blackstock
Keyword(s):  
Chronic Obstructive Pulmonary Disease ◽  
Literature Review ◽  
Functional Status ◽  
Daily Activity ◽  
Psychometric Testing ◽  
Chronic Obstructive ◽  
Activity Levels ◽  
Critical Feature ◽  
Obstructive Pulmonary Disease ◽  
Categorical Framework

The objective of this study is to review available functional status measures (FSMs) validated for use in the chronic obstructive pulmonary disease (COPD) population and categorizing the measures by their commonalities to formulate a framework that supports clinicians in the selection and application of FSMs. A literature review identifying valid and reliable measures of functional status for people with COPD was undertaken. Measures were thematically analyzed and categorized to develop a framework for clinical application. A variety of measures of activity levels exist, with 35 included in this review. Thematic categorization identified five categories of measures: daily activity, impact, surrogate, performance-based, and disability-based measures. The vast variety of FSMs available for clinicians to apply with people who have COPD may be overwhelming, and selection must be thoughtfully based on the nature of the population being studied/evaluated, and aims of evaluation being conducted, not simply as a standard measure used at the institution. Psychometric testing is a critical feature to a strong instrument and issues of reliability, validity, and responsiveness need to be understood prior to measurement use. Contextual nature of measures such as language used and activities measured is also important. A categorical framework to support clinicians in the selection and application of FSMs has been presented in this article.

scholarly journals Agrobacterium-Mediated Gene Transient Overexpression and Tobacco Rattle Virus (TRV)-Based Gene Silencing in Cassava

2019 ◽  
Vol 20 (16) ◽  
pp. 3976 ◽  
Author(s):  
Hongqiu Zeng ◽  
Yanwei Xie ◽  
Guoyin Liu ◽  
Yunxie Wei ◽  
Wei Hu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Functional Genomics ◽  
Transient Expression ◽  
Fluorescent Protein ◽  
Tobacco Rattle Virus ◽  
Mosaic Disease ◽  
African Cassava Mosaic Virus ◽  
Cassava Mosaic Disease ◽  
Virus Induced Gene Silencing ◽  
Transient Expression Assay

Agrobacterium-mediated transient expression and virus-induced gene silencing (VIGS) are very useful in functional genomics in plants. However, whether these methods are effective in cassava (Manihot esculenta), one of the most important tropical crops, remains elusive. In this study, we used green fluorescent protein (GFP) and β-glucuronidase (GUS) as reporter genes in a transient expression assay. GFP or GUS could be detected in the infiltrated leaves at 2 days postinfiltration (dpi) and were evidenced by visual GFP and GUS assays, reverse-transcription PCR, and Western blot. In addition, phytoene desaturase (PDS) was used to show the silencing effect in a VIGS system. Both Agrobacterium GV3101 and AGL-1 with tobacco rattle virus (TRV)-MePDS-infiltrated distal leaves showed an albino phenotype at 20 dpi; in particular, the AGL-1-infiltrated plants showed an obvious albino area in the most distal leaves. Moreover, the silencing effect was validated by molecular identification. Notably, compared with the obvious cassava mosaic disease symptom infiltrated by African-cassava-mosaic-virus-based VIGS systems in previous studies, TRV-based VIGS-system-infiltrated cassava plants did not show obvious virus-induced disease symptoms, suggesting a significant advantage. Taken together, these methods could promote functional genomics in cassava.

scholarly journals Transfected human beta-polymerase promoter contains a ras-responsive element.

1990 ◽  
Vol 10 (7) ◽  
pp. 3852-3856 ◽  
Author(s):  
P S Kedar ◽  
D R Lowy ◽  
S G Widen ◽  
S H Wilson
Keyword(s):  
Dna Repair ◽  
Transient Expression ◽  
Dna Polymerase ◽  
Genomic Dna ◽  
3T3 Cells ◽  
Gap Filling ◽  
Transient Expression Assay ◽  
Responsive Element ◽  
Cellular Dna ◽  
Nih 3T3

beta-Polymerase is a vertebrate cellular DNA polymerase involved in gap-filling synthesis during some types of genomic DNA repair. We report that a cloned human beta-polymerase promoter in a transient expression assay is activated by p21v-rasH expression in NIH 3T3 cells. A decanucleotide palindromic element, GTGACGTCAC, at positions -49 to -40 in the promoter is required for this ras-mediated stimulation.

Concentration of bat activity in riparian habitats over an elevational gradient

1999 ◽  
Vol 77 (6) ◽  
pp. 972-977 ◽  
Author(s):  
S D Grindal ◽  
J L Morissette ◽  
R M Brigham
Keyword(s):  
Elevational Gradient ◽  
Relative Activity ◽  
Activity Levels ◽  
Riparian Areas ◽  
Habitat Types ◽  
Bat Activity ◽  
Prey Resources ◽  
Riparian Habitats ◽  
Mist Net ◽  
Capture Rates

Riparian areas are generally assumed to represent important foraging areas for insectivorous bats, but this contention has rarely been formally quantified. To test this assumption, we used bat detectors to compare the relative activity levels of a community of temperate-zone bat species between riparian (lake) and upland (cutblock) habitats at three different elevations (ranging from 540 to 1800 m) in a forested area of southern British Columbia. In addition, we also investigated the sex and age class distributions of bats (based on mist-net captures) between riparian and upland habitats among the elevational zones. Bat activity levels were significantly greater in riparian than upland areas (10 and 40 times greater for foraging and commuting activity, respectively). Capture rates were greater in riparian areas and biased towards females, suggesting that female bats may preferentially select riparian areas, probably because of the abundant prey resources typically associated with this habitat. Captures of females also predominated at lower elevations, whereas males were captured more often in higher elevation zones. Our data support the assumption that riparian habitats represent important foraging and probably drinking areas for bats. The sex bias and differences in capture rates and activity levels need to be considered when designing bat surveys in different habitat types or over elevational gradients.

Transient expression assay for qualitative assessment of gene expression by fowlpox virus

Virus Genes ◽  
1990 ◽  
Vol 3 (3) ◽  
pp. 213-220 ◽  
Author(s):  
Shree Dhawale ◽  
Christopher E. Beisel ◽  
Keyvan Nazerian
Keyword(s):  
Gene Expression ◽  
Transient Expression ◽  
Qualitative Assessment ◽  
Transient Expression Assay ◽  
Fowlpox Virus

Characteristics of chalcone isomerase promoter in crabapple leaves (M.cv. royalty) and transient expression assay modified in onion epidermal cell

2011 ◽  
Vol 10 (50) ◽  
pp. 10232-10240
Author(s):  
Tian Ji ◽  
Shen Hongxiang ◽  
Zhang Jie ◽  
Song Tingting ◽  
Yao Yuncong
Keyword(s):  
Transient Expression ◽  
Epidermal Cell ◽  
Chalcone Isomerase ◽  
Transient Expression Assay ◽  
Onion Epidermal Cell

scholarly journals Transient Expression Assay in Strawberry Fruits

BIO-PROTOCOL ◽  
2019 ◽  
Vol 9 (11) ◽  
Author(s):  
Mengting Pi ◽  
Qi Gao ◽  
Chunying Kang
Keyword(s):  
Transient Expression ◽  
Transient Expression Assay

scholarly journals Activation of the Transcription of BrGA20ox3 by a BrTCP21 Transcription Factor Is Associated with Gibberellin-Delayed Leaf Senescence in Chinese Flowering Cabbage during Storage

2019 ◽  
Vol 20 (16) ◽  
pp. 3860
Author(s):  
Xian-mei Xiao ◽  
Yan-mei Xu ◽  
Ze-xiang Zeng ◽  
Xiao-li Tan ◽  
Zong-li Liu ◽  
...  
Keyword(s):  
Transient Expression ◽  
Leaf Senescence ◽  
Biosynthetic Pathway ◽  
Transcript Level ◽  
Mobility Shift ◽  
Transient Expression Assay ◽  
Catabolic Genes ◽  
Mobility Shift Assay ◽  
Psii Quantum Yield ◽  
Senescence Associated Genes

Several lines of evidence have implicated the involvement of the phytohormone gibberellin (GA) in modulating leaf senescence in plants. However, upstream transcription factors (TFs) that regulate GA biosynthesis in association with GA-mediated leaf senescence remain elusive. In the current study, we report the possible involvement of a TEOSINTE BRANCHED1/CYCLOIDEA/PCF (TCP) TF BrTCP21 in GA-delayed leaf senescence in Chinese flowering cabbage. Exogenous GA3 treatment maintained a higher value of maximum PSII quantum yield (Fv/Fm) and total chlorophyll content, accompanied by the repression of the expression of senescence-associated genes and chlorophyll catabolic genes, which led to the delay of leaf senescence. A class I member of TCP TFs BrTCP21, was further isolated and characterized. The transcript level of BrTCP21 was low in senescing leaves, and decreased following leaf senescence, while GA3 could keep a higher expression level of BrTCP21. BrTCP21 was further found to be a nuclear protein and exhibit trans-activation ability through transient-expression analysis in tobacco leaves. Intriguingly, the electrophoretic mobility shift assay (EMSA) and transient expression assay illustrated that BrTCP21 bound to the promoter region of a GA biosynthetic gene BrGA20ox3, and activated its transcription. Collectively, these observations reveal that BrTCP21 is associated with GA-delayed leaf senescence, at least partly through the activation of the GA biosynthetic pathway. These findings expand our knowledge on the transcriptional mechanism of GA-mediated leaf senescence.

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