A Human cDNA Library for High-Throughput Protein Expression Screening

Genomics ◽  
2000 ◽  
Vol 65 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Konrad Büssow ◽  
Eckhard Nordhoff ◽  
Christine Lübbert ◽  
Hans Lehrach ◽  
Gerald Walter
Keyword(s):  
Protein Expression ◽  
Cdna Library ◽  
High Throughput ◽  
Expression Screening ◽  
Human Cdna

High-throughput protein expression screening and purification in Escherichia coli

Methods ◽  
2011 ◽  
Vol 55 (1) ◽  
pp. 65-72 ◽  
Author(s):  
Renaud Vincentelli ◽  
Agnès Cimino ◽  
Arie Geerlof ◽  
Atsushi Kubo ◽  
Yutaka Satou ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein Expression ◽  
High Throughput ◽  
Expression Screening

scholarly journals High Throughput Screen Identifies the DNMT1 (DNA Methyltransferase-1) Inhibitor, 5-Azacytidine, as a Potent Inducer of PTEN (Phosphatase and Tensin Homolog)

2020 ◽  
Vol 40 (8) ◽  
pp. 1854-1869
Author(s):  
Keith A. Strand ◽  
Sizhao Lu ◽  
Marie F. Mutryn ◽  
Linfeng Li ◽  
Qiong Zhou ◽  
...  
Keyword(s):  
Protein Expression ◽  
High Throughput ◽  
Knockout Mice ◽  
Vascular Remodeling ◽  
Dna Methyltransferase ◽  
Dna Methyltransferase 1 ◽  
Protective Effects ◽  
High Throughput Screen ◽  
Phosphatase And Tensin Homolog ◽  
Tensin Homolog

Objective: Our recent work demonstrates that PTEN (phosphatase and tensin homolog) is an important regulator of smooth muscle cell (SMC) phenotype. SMC-specific PTEN deletion promotes spontaneous vascular remodeling and PTEN loss correlates with increased atherosclerotic lesion severity in human coronary arteries. In mice, PTEN overexpression reduces plaque area and preserves SMC contractile protein expression in atherosclerosis and blunts Ang II (angiotensin II)-induced pathological vascular remodeling, suggesting that pharmacological PTEN upregulation could be a novel therapeutic approach to treat vascular disease. Approach and Results: To identify novel PTEN activators, we conducted a high-throughput screen using a fluorescence based PTEN promoter-reporter assay. After screening ≈3400 compounds, 11 hit compounds were chosen based on level of activity and mechanism of action. Following in vitro confirmation, we focused on 5-azacytidine, a DNMT1 (DNA methyltransferase-1) inhibitor, for further analysis. In addition to PTEN upregulation, 5-azacytidine treatment increased expression of genes associated with a differentiated SMC phenotype. 5-Azacytidine treatment also maintained contractile gene expression and reduced inflammatory cytokine expression after PDGF (platelet-derived growth factor) stimulation, suggesting 5-azacytidine blocks PDGF-induced SMC de-differentiation. However, these protective effects were lost in PTEN-deficient SMCs. These findings were confirmed in vivo using carotid ligation in SMC-specific PTEN knockout mice treated with 5-azacytidine. In wild type controls, 5-azacytidine reduced neointimal formation and inflammation while maintaining contractile protein expression. In contrast, 5-azacytidine was ineffective in PTEN knockout mice, indicating that the protective effects of 5-azacytidine are mediated through SMC PTEN upregulation. Conclusions: Our data indicates 5-azacytidine upregulates PTEN expression in SMCs, promoting maintenance of SMC differentiation and reducing pathological vascular remodeling in a PTEN-dependent manner.

scholarly journals High throughput transformation of a Sorghum cDNA library for rice improvement

2016 ◽  
Vol 125 (3) ◽  
pp. 471-478 ◽  
Author(s):  
Kezhou Qin ◽  
Ping Qiu ◽  
Jianyu Wen ◽  
Yingguo Zhu ◽  
Nengwu Li ◽  
...  
Keyword(s):  
Cdna Library ◽  
High Throughput ◽  
Rice Improvement
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