Gene expression associated with light-induced conidiation in Colletotrichum graminicola

1991 ◽  
Vol 37 (2) ◽  
pp. 165-167 ◽  
Author(s):  
Zhenfan Yang ◽  
Daniel G. Panaccione ◽  
Robert M. Hanau
Keyword(s):  
Gene Expression ◽  
Key Words ◽  
Agar Medium ◽  
Cell Development ◽  
Conidiogenous Cell ◽  
In Vitro Translation ◽  
Asexual Development ◽  
Light Induction ◽  
Colletotrichum Graminicola

Light was used to induce conidiation in uniform populations of vegetative hyphae of Collectotrichum graminicola grown on agar medium. Differentiation of conidiogenous cells, the first detectable event in conidial morphogenesis, was rapid and highly synchronized. In vitro translation of poly(A)+ RNA from dark-grown (nondifferentiating) and light-induced (differentiating) hyphae demonstrated that conidiogenous cell development was accompanied by detectable changes in gene expression. Key words: Colletotrichum, conidia, asexual development, conidiogenous cell, light induction.

scholarly journals Dynamic regulation of chromatin organizer SATB1 via TCR-induced alternative promoter switch during T-cell development

2020 ◽  
Vol 48 (11) ◽  
pp. 5873-5890
Author(s):  
Indumathi Patta ◽  
Ayush Madhok ◽  
Satyajeet Khare ◽  
Kamalvishnu P Gottimukkala ◽  
Anjali Verma ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Development ◽  
Cell Development ◽  
Alternative Promoter ◽  
In Vitro Translation ◽  
Alternative Promoters ◽  
Transcript Variants ◽  
P2 Promoter ◽  
Specific Regulation

Abstract The chromatin organizer SATB1 is highly enriched in thymocytes and is essential for T-cell development. Although SATB1 regulates a large number of genes important for T-cell development, the mechanism(s) regulating expression of SATB1 during this process remain elusive. Using chromatin immune precipitation-seq-based occupancy profiles of H3K4me3 and H3Kme1 at Satb1 gene locus, we predicted four different alternative promoters of Satb1 in mouse thymocytes and characterized them. The expression of Satb1 transcript variants with distinct 5′ UTRs occurs in a stage-specific manner during T-cell development and is dependent on TCR signaling. The observed discrepancy between the expression levels of SATB1 mRNA and protein in developing thymocytes can be explained by the differential translatability of Satb1 transcript variants as confirmed by polysome profiling and in vitro translation assay. We show that Satb1 alternative promoters exhibit lineage-specific chromatin accessibility during T-cell development from progenitors. Furthermore, TCF1 regulates the Satb1 P2 promoter switch during CD4SP development, via direct binding to the Satb1 P2 promoter. CD4SP T cells from TCF1 KO mice exhibit downregulation of P2 transcript variant expression as well as low levels of SATB1 protein. Collectively, these results provide unequivocal evidence toward alternative promoter switch-mediated developmental stage-specific regulation of SATB1 in thymocytes.

scholarly journals Blockage of tropoelastin secretion by monensin represses tropoelastin synthesis at a pretranslational level in rat smooth muscle cells.

1985 ◽  
Vol 5 (1) ◽  
pp. 253-258 ◽  
Author(s):  
S M Frisch ◽  
J M Davidson ◽  
Z Werb
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Genomic Dna ◽  
Mrna Level ◽  
In Vitro Translation ◽  
Aortic Smooth Muscle Cell ◽  
Aortic Smooth Muscle ◽  
Reported Effect ◽  
Ca 50

The blockage of protein secretion in the R22 cultured rat aortic smooth muscle cell strain with monensin repressed tropoelastin gene expression at the mRNA level by ca. 50-fold as measured by biosynthetic pulse-labeling, in vitro translation, and hybridization with a tropoelastin genomic DNA probe. These results suggest that tropoelastin gene expression is autoregulated, and they represent the first reported effect of monensin on gene expression.

Evaluation of Role for flt3L in Human Early B and T/NK Lymphopoiesis in a Novel Stromal Cell Culture System

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 2385-2385
Author(s):  
Yoshiki Nakamori ◽  
Kohshi Ohishi ◽  
Bing Liu ◽  
Masahiro Masuya ◽  
Hirofumi Hamada ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stromal Cells ◽  
Culture Condition ◽  
Culture System ◽  
Nk Cell ◽  
Cell Development ◽  
Cellular Interaction ◽  
Differentiation Potential ◽  
Vitro Assay

Abstract Abstract 2385 The regulatory mechanism of human early lymphopoiesis remains less defined. A major limitation of conventional in vitro assay is that B and T lymphopoiesis cannot be assessed simultaneously. We exploded a novel culture assay and found that the hTERT-transduced telomerized human stromal cells support the generation of CD19+CD34lo/-CD10+cyCD79a+CD20+/−VpreB− pro B- and CD7+CD34+CD45RA+CD56−cyCD3− early T-cell precursors from human hematopoietic progenitors without cytokines, which was enhanced by flt3L (2010, ASH). We further characterized the generated lymphoid precursors, and verified that lineage-specific transcription factors for B (Pax-5 and EBF) and T/NK-cell precursors (GATA-3, HEB, Id2) were expressed by the CD19+ and CD7+CD56− cells, respectively. The CD7+CD56− cells showed the differentiation potential to T- and NK-lineage cells, by replating with OP9-Delta1 in the presence of flt3L+IL-7, and with the telomerized-stromal cells in the presence of flt3L+IL-7+IL-15, respectively. In serum-free culture condition, B-cell differentiation was minimally supported by the stromal cells, while low number of CD7+ cells was observed. Nonetheless, a significant number of CD7+, CD19-cyCD79a+, and CD19+ cells developed in the presence of flt3L, suggesting an important role for flt3L in the generation of early T/NK- and, particularly, B-lineage cell precursors on the stromal cells. The cellular interaction between Notch and Notch ligand Delta-1 or -4 in the presence of appropriate cytokines is considered to be crucial in T-lineage cell development in mice. We therefore examined whether Notch pathway is involved in the T/NK-cell development under this culture condition. While the gene expression of Delta-1 or -4 was detected in the telomerized stromal cells, the protein expression of these ligands was not detected with Western-blotting analysis. After co-culture of hematopoietic progenitor cells with the telomerized stromal cells, the gene expression of Notch target gene, HES1, was not increased. These data suggest that Notch signaling is not involved in the generation of early T/NK cell precursors on the stromal cells. This novel in vitro culture system suggests that the development of early B- and T/NK- cell precursors from hematopoietic precursors take places on the stromal cells in a Notch-independent manner and that flt3L plays a principal role in the stimulation of the early B and T/NK lymphopoiesis. Disclosures: No relevant conflicts of interest to declare.

scholarly journals KARAKTERISTIK FISIOLOGI ISOLAT Pleurotus spp.

2020 ◽  
Vol 15 (1) ◽  
pp. 46
Author(s):  
ACHMAD ACHMAD ◽  
ELIS NINA HERLIYANA ◽  
OSICA ASNO FERINA YURTI ◽  
ANANG PRANOTO HIDAYAT
Keyword(s):  
Key Words ◽  
Gallic Acid ◽  
Agar Medium ◽  
Physiological Characteristics ◽  
Medium Temperature ◽  
Physiological Character ◽  
Diameter Range ◽  
Temperature Levels

<p>ABSTRAK</p><p>Studi in vitro tentang karakteristik fungi isolat Pleurotus spp. telahdilaksanakan di Bogor dari bulan Juli sampai Agustus 2004. Penelitianmenggunakan rancangan faktorial dalam rancangan acak lengkap danbertujuan untuk mempelajari pengaruh media, temperatur inkubasi dan pHmedia terhadap 6 isolat Pleurotus sp. Karakter lain yang juga dipelajariadalah kemampuan untuk mengoksidasi asam tanat dan asam galat dalammedia agar. Hasil penelitian menunjukkan bahwa Pleurotus isolat sp.6dan sp.8 tumbuh baik pada media MPA, isolat Pleurotus sp.1, 3 dan 4pada media MEA dan isolate sp.2 pada media PDA. Kecuali isolat sp.8,isolat lainnya tidak dapat tumbuh pada temperatur 10 dan 35 o C. Pertum-buhan isolat sp.8 terbaik dibandingkan isolat lainnya pada semuatemperatur. Diameter koloni isolat lainnya hanya mencapai 0,2 – 2,33 cm.Pertumbuhan isolat sp.8 juga terbaik pada semua pH media diikuti isolatsp.6 kemudian isolat sp.4. Semua isolat menunjukkan reaksi oksidasipositif pada agar asam tanat dan asam galat yang ditunjukkan oleh warnacoklat pada media yang melingkari koloni.</p><p>Kata kunci : Pleurotus spp, media, temperature, pH, oksidasi, asam tanat,asam galat</p><p>ABSTRACT</p><p>Physiological Characteristics of Pleurotus spp. IsolatesPhysiological characteristics of some Pleurotus sp. isolates werestudied in vitro, from July until August 2004 in Bogor. Experiments tostudy the effect of kind of media, temperature of incubation room, and pHof medium on six isolates of Pleurotus sp. were arranged in factorialrandomized complete design and replicated three times with colony in apetri dish as experimental units. Another physiological character studiedwas the ability to oxidize tannic and gallic acids in agar medium. Resultsshowed that isolates Pleurotus sp.6 and -8 grew better in MPA medium,Pleurotus sp.1, -3, and -4 in MEA, and Pleurotus sp.2 in PDA. ExceptPleurotus sp.8, other isolates could not grow in incubation roomtemperature of 10 and 35 o  C. The growth of Pleurotus sp.8 was the bestamong the isolates in all temperature levels. Other isolates grew poorly in20 and 29o C with diameter range was 0.2 – 2.33 cm. The growth ofPleurotus sp.8 was also the best in all pH medium levels, followed byPleurotus sp.6, and then Pleurotus sp.4. All isolates showed positiveoxidative reaction on tannic and gallic acid agar indicated by brown colorof the medium around the colony.</p><p>Key words: Pleurotus spp., medium, temperature, pH, oxidation, tannicacid, gallic acid</p>

scholarly journals Production of human translation-competent lysates using dual centrifugation

2021 ◽  
Author(s):  
Lukas-Adrian Gurzeler ◽  
Jana Ziegelmuller ◽  
Oliver Muhlemann ◽  
Evangelos D. Karousis
Keyword(s):  
Gene Expression ◽  
Cell Lysis ◽  
Human Cells ◽  
In Vitro Translation ◽  
Fast Method ◽  
Central Process ◽  
Translation Systems

Translation is a central process in gene expression and the development of efficient in vitro translation systems has been the focus of scientific efforts for many decades. The production of translation-competent lysates originating from human cells or tissues remains challenging, mainly due to the variability of cell lysis conditions. Here we present a robust and fast method based on dual centrifugation that allows the preparation of cytoplasmic extracts from human cells that efficiently translate mRNAs in a capped and IRES-mediated way. We optimized lysate preparation and in vitro translation conditions and show that dual centrifugation allows the production of human lysates under detergent-free conditions and is ideally suited to produce ample amounts of human translation-competent lysates.

Blockage of tropoelastin secretion by monensin represses tropoelastin synthesis at a pretranslational level in rat smooth muscle cells

1985 ◽  
Vol 5 (1) ◽  
pp. 253-258
Author(s):  
S M Frisch ◽  
J M Davidson ◽  
Z Werb
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Genomic Dna ◽  
Mrna Level ◽  
In Vitro Translation ◽  
Aortic Smooth Muscle Cell ◽  
Aortic Smooth Muscle ◽  
Reported Effect ◽  
Ca 50

The blockage of protein secretion in the R22 cultured rat aortic smooth muscle cell strain with monensin repressed tropoelastin gene expression at the mRNA level by ca. 50-fold as measured by biosynthetic pulse-labeling, in vitro translation, and hybridization with a tropoelastin genomic DNA probe. These results suggest that tropoelastin gene expression is autoregulated, and they represent the first reported effect of monensin on gene expression.

Enforced Expression of Pax-5 Results in Developmental Arrest, Immortalisation and Aberrant Expression of B Lineage Genes in Committed Myeloid Progenitors.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2784-2784
Author(s):  
Kristina Anderson ◽  
Corinne Rusterholz ◽  
Robert Mansson ◽  
Christina Jensen ◽  
Karl Bacos ◽  
...  
Keyword(s):  
Gene Expression ◽  
B Cell ◽  
Gene Expression Pattern ◽  
Cell Development ◽  
B Cell Development ◽  
Myeloid Differentiation ◽  
Aberrant Expression ◽  
Murine Bone Marrow ◽  
Myeloid Progenitors

Abstract The paired domain transcription factor Pax-5, has been demonstrated to play a crucial role in development and final commitment of the B cell lineage. In fact, otherwise committed B cell progenitors acquire multipotentiality (myelo-lymphoid) potential upon targeted deletion of Pax-5 expression (Nutt et al. Nature.1999; 401:556–562). Thus, in addition, to promoting B cell development through acting as an activator of transcription of B cell specific genes such as CD19, Pax-5 is also thought to act as a suppressor of transcription of genes involved in determination of other blood cell lineages. However, it remains unclear how Pax-5 might repress myeloid development. Thus, we investigated the effect of Pax-5 expression on lympho-myeloid differentiation by overexpressing human (h)Pax-5 (through retroviral transduction) in adult murine bone marrow Linlo/−Sca-1+c-kit+ (LSK) cells. When compared to cells transduced with a control vector, LSK cells ectopically expressing hPax-5 very efficiently developed into hPax-5+B220+CD19+ pro-B cells in response to flt3 ligand and interleukin-7 in vitro. In contrast, when hPax-5+ LSK cells were cultured under myeloid conditions, we consistently observed development of a highly proliferative and immortalised bi-phenotypic (B-myeloid) hPax-5+B220+Gr-1+Mac-1+ population that predominantly consisted of immature myeloblasts but also maturated granulocytes and monocytes. Global gene expression analysis by micro-array, and confirmation by RT-PCR, demonstrated that hPax-5+B220+Gr-1+Mac-1+ cells also possessed a bi-phenotypic gene expression pattern, characteristic for B-cell as well as myeloid lineages including the Pax-5 target B cell genes mb-1 and BLNK as well as GM-CSFRα and low levels of C/EBPα. Similar findings were observed when targeting committed myeloid progenitors. These findings suggest that in addition to promoting B cell development, Pax-5 is capable of inhibiting/blocking myeloid differentiation and inducing immortalization as well as expression of B cell specific genes in otherwise myeloid committed progenitors. These findings motivate a careful investigation of the potential involvement of Pax-5 also in myeloid leukemias.

In vitro translation in a hybrid cell free lysate with exogenous cellular ribosomes

2015 ◽  
Vol 467 (3) ◽  
pp. 387-398 ◽  
Author(s):  
Baptiste Panthu ◽  
Didier Décimo ◽  
Laurent Balvay ◽  
Théophile Ohlmann
Keyword(s):  
Gene Expression ◽  
Protein Synthesis ◽  
Hybrid System ◽  
Hybrid Cell ◽  
Cell Types ◽  
In Vitro Translation ◽  
Translation System ◽  
Different Origins ◽  
Cell Free Protein Synthesis

Cell free protein synthesis systems (CFPS) have been widely used to express proteins and to explore the pathways of gene expression. In the present manuscript, we describe the design of a novel adaptable hybrid in vitro translation system which is assembled with ribosomes isolated from many different origins. We first show that this hybrid system exhibits all important features such as efficiency, sensitivity, reproducibility and the ability to translate specialized mRNAs in less than 1 h. In addition, the unique design of this cell free assay makes it highly adaptable to utilize ribosomes isolated from many different organs, tissues or cell types.

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