High-level secretion of Pseudomonas fluorescens type I secretion system-dependent lipase in Serratia marcescens

2007 ◽  
Vol 130 (3) ◽  
pp. 311-315 ◽  
Author(s):  
Jae Kwang Song ◽  
Joon Young Oh ◽  
Gyeong Tae Eom ◽  
Bong Keun Song
Keyword(s):  
Serratia Marcescens ◽  
Pseudomonas Fluorescens ◽  
Secretion System ◽  
Type I ◽  
High Level

scholarly journals Utilizing the ABC Transporter for Growth Factor Production by fleQ Deletion Mutant of Pseudomonas fluorescens

Biomedicines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 679
Author(s):  
Benedict-Uy Fabia ◽  
Joshua Bingwa ◽  
Jiyeon Park ◽  
Nguyen-Mihn Hieu ◽  
Jung-Hoon Ahn
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Pseudomonas Fluorescens ◽  
Abc Transporter ◽  
Transforming Growth Factor Beta ◽  
Deletion Mutant ◽  
Transforming Growth Factor ◽  
Gene Knockout ◽  
Type I ◽  
Double Deletion

Pseudomonas fluorescens, a gram-negative bacterium, has been proven to be a capable protein manufacturing factory (PMF). Utilizing its ATP-binding cassette (ABC) transporter, a type I secretion system, P. fluorescens has successfully produced recombinant proteins. However, besides the target proteins, P. fluorescens also secretes unnecessary background proteins that complicate protein purification and other downstream processes. One of the background proteins produced in large amounts is FliC, a flagellin protein. In this study, the master regulator of flagella gene expression, fleQ, was deleted from P. fluorescens Δtp, a lipase and protease double-deletion mutant, via targeted gene knockout. FleQ directs flagella synthesis, so the new strain, P. fluorescens ΔfleQ, does not produce flagella-related proteins. This not only simplifies purification but also makes P. fluorescens ΔfleQ an eco-friendly expression host because it will not survive outside a controlled environment. Six recombinant growth factors, namely, insulin-like growth factors I and II, beta-nerve growth factor, fibroblast growth factor 1, transforming growth factor beta, and tumor necrosis factor beta, prepared using our supercharging method, were successfully secreted by P. fluorescens ΔfleQ. Our findings demonstrate the potential of P. fluorescens ΔfleQ, combined with our supercharging process, as a PMF.

Experiments with a biological material for the closure of incisional hernias

1987 ◽  
Vol 21 (3) ◽  
pp. 195-200 ◽  
Author(s):  
M. Walter ◽  
U. Brenner ◽  
W. Holzmüller ◽  
J. M. Müller
Keyword(s):  
Collagen Type I ◽  
Morphological Properties ◽  
Type I ◽  
Native Structure ◽  
Incisional Hernias ◽  
Small Vessels ◽  
Collagen Implant ◽  
Infiltrating Cells ◽  
High Level ◽  
Special Case

A new preparation process was studied which should allow the implantation of collagen type I in its native structure in reconstructive surgery, in this special case for closure of incisional hernias. As experimental animals we used 30 female Lewis rats. A defect of the anterior abdominal wall measuring 3 cm × 4 cm was closed with our collagen substitute. Biopsies taken after 4, 6 and 8 weeks were examined morphologically. As criteria for revitalization and revascularization we used the type of infiltrating cells, the depth and density of infiltration and the formation of new blood vessels. After 4 weeks the implants were infiltrated by fibroblasts that decreased in density towards the centre. Good revascularization could be seen on the muscle-implant interface. After 6 weeks the density of infiltrating cells had increased markedly even to the centre of the collagen implant. Sporadically small vessels could be seen. Eight weeks after implantation the density of infiltrated cells was at the same high level, and capillary bundles could be seen within the whole implant. We believe that this collagen implant is suitable for the closure of hernias as shown by its physical and morphological properties. In particular it appears to guarantee an earlier and tighter closure of hernias than other materials.

scholarly journals An A/U-Rich Enhancer Region Is Required for High-Level Protein Secretion through the HlyA Type I Secretion System

2017 ◽  
Vol 84 (1) ◽  
Author(s):  
Sakshi Khosa ◽  
Romy Scholz ◽  
Christian Schwarz ◽  
Mirko Trilling ◽  
Hartmut Hengel ◽  
...  
Keyword(s):  
Ribosomal Protein ◽  
Protein Secretion ◽  
Fusion Proteins ◽  
Untranslated Region ◽  
Secretion System ◽  
Type I ◽  
Secretion Systems ◽  
Enhancer Region ◽  
Ribosomal Protein S1 ◽  
Cellular Expression

ABSTRACTEfficient protein secretion is often a valuable alternative to classic cellular expression to obtain homogenous protein samples. Early on, bacterial type I secretion systems (T1SS) were employed to allow heterologous secretion of fusion proteins. However, this approach was not fully exploited, as many proteins could not be secreted at all or only at low levels. Here, we present an engineered microbial secretion system which allows the effective production of proteins up to a molecular mass of 88 kDa. This system is based on the hemolysin A (HlyA) T1SS of the Gram-negative bacteriumEscherichia coli, which exports polypeptides when fused to a hemolysin secretion signal. We identified an A/U-rich enhancer region upstream ofhlyArequired for effective expression and secretion of selected heterologous proteins irrespective of their prokaryotic, viral, or eukaryotic origin. We further demonstrate that the ribosomal protein S1 binds to thehlyAA/U-rich enhancer region and that this region is involved in the high yields of secretion of functional proteins, like maltose-binding protein or human interferon alpha-2.IMPORTANCEA 5′ untranslated region of the mRNA of substrates of type I secretion systems (T1SS) drastically enhanced the secretion efficiency of the endogenously secreted protein. The identification of ribosomal protein S1 as the interaction partner of this 5′ untranslated region provides a rationale for the enhancement. This strategy furthermore can be transferred to fusion proteins allowing a broader, and eventually a more general, application of this system for secreting heterologous fusion proteins.

scholarly journals Citrullinemia Type I - A Case Report

2016 ◽  
Vol 33 (2) ◽  
pp. 98-100
Author(s):  
Fauzia Mohsin ◽  
Sharmin Mahbuba ◽  
Tahmina Begum ◽  
Narayan Chandra Saha ◽  
Kishwar Azad ◽  
...  
Keyword(s):  
Urea Cycle ◽  
Female Child ◽  
Amino Acid Profile ◽  
Dramatic Improvement ◽  
Type I ◽  
Frequent Vomiting ◽  
Cycle Disorders ◽  
One Year ◽  
High Level ◽  
Citrullinemia Type I

Citrullinemia type I (CTLN1) is an inherited urea cycle disorder where the enzyme argininosuccinate synthetase is deficient. It can lead to recurrent hyperammonemic crisis that may result in permanent neurological sequelae, even death. Vomiting in patients with urea cycle disorders may either be the result or cause of acute hyperammonemia, particularly if due to an illness that leads to catabolism. Therefore, age-appropriate common etiologies of vomiting must be considered when evaluating these patients. We present a case of a 2 year 5 month old female child with CTLN1 who had a history of frequent vomiting after the age of one year and some recent neurological manifestations like excessive crying and lethargy and one episode of unconsciousness. Investigations revealed high level of ammonia. Amino acid profile using tandem mass spectrometry showed markedly increased plasma level of citrulline. After administration of sodium benzoate and protein restricted diet there was dramatic improvement of all the symptoms.J Bangladesh Coll Phys Surg 2015; 33(2): 98-100

scholarly journals Rizobacterias con potencial antagonista in vitro a Mycosphaerella fijiensis Morelet

2020 ◽  
Vol 13 (2) ◽  
Author(s):  
Karen Tatiana Chávez Arteaga ◽  
Jefferson Javier Guato Molina ◽  
Jorge Luis Rodríguez Acosta ◽  
Ángel Virgilio Cedeño Moreira ◽  
Ricardo Fernando Romero Meza ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Pseudomonas Putida ◽  
Serratia Marcescens ◽  
Pseudomonas Fluorescens ◽  
Mycosphaerella Fijiensis ◽  
Subtilis Atcc ◽  
Bacillus Subtilis Atcc ◽  
Pseudomonas Veronii ◽  
Enterobacter Asburiae

El empleo de bio-controladores en la agricultura beneficia los aspectos fisiológicos en plantas, a diferencia de la constante aplicación de pesticidas en el cultivo del banano ha ocasionado la pérdida de la sensibilidad en M. fijiensis, reduciendo la microbiota del suelo. El objetivo se enfocó en caracterizar el potencial antagónico de las PGPR en inhibición de germinación de ascósporas y desarrollo micelial de M. fijiensis. Se realizaron cultivos monospóricos de M. fijiensis e identificado por PCR. Se evaluaron los extractos celulares de Pseudomonas putida PB3-6, Klebsiella variicola BO3-4, Enterobacter asburiae BA4-19, Serratia marcescens PM3-8, Enterobacter asburiae PM3-14, Pseudomonas protegens CHA0, Pseudomonas fluorescens WCS417, Pseudomonas veronii R4 y Bacillus subtilis ATCC 5540 para sus evaluaciones antagonistas: a) Inhibición del tubo germinativo de las ascósporas al 2% y b) Desarrollo micelial al (2 y 10 %). La PCR empleado en la identificación de M. fijiensis se confirma el producto de amplificación de 1018 pb. El factor antagónico de los extractos celulares al 2 % de PM3-14 y CHA0 inhibe sobre el 80 % al desarrollo de los tubos germinativos. La inhibición al desarrollo micelial del extracto celular al 2 %, de CHA0 logró una efectividad del 54 % y las cepas (PM3-8, PM3-14 y BA4-19) con (32, 26 y 26 %). Al 10 % del extracto de la cepa PM3-8 inhibe el desarrollo micelial con niveles de turbidez de 0,47 (OD600nm). El empleo de estos bio-controladores en la agricultura ofrecerá una alternativa para beneficiar en la reducción del uso de agroquímicos

Pseudomonas fluorescens C7R12 type III secretion system impacts mycorrhization of Medicago truncatula and associated microbial communities

Mycorrhiza ◽  
2016 ◽  
Vol 27 (1) ◽  
pp. 23-33 ◽  
Author(s):  
Amandine Viollet ◽  
Barbara Pivato ◽  
Christophe Mougel ◽  
Jean-Claude Cleyet-Marel ◽  
Cécile Gubry-Rangin ◽  
...  
Keyword(s):  
Microbial Communities ◽  
Pseudomonas Fluorescens ◽  
Medicago Truncatula ◽  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Type Iii

scholarly journals High-level transgene expression in induced pluripotent stem cell–derived megakaryocytes: correction of Glanzmann thrombasthenia

Blood ◽  
2014 ◽  
Vol 123 (5) ◽  
pp. 753-757 ◽  
Author(s):  
Spencer K. Sullivan ◽  
Jason A. Mills ◽  
Sevasti B. Koukouritaki ◽  
Karen K. Vo ◽  
Randolph B. Lyde ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Type I ◽  
Glanzmann Thrombasthenia ◽  
Key Points ◽  
Single Allele ◽  
Induced Pluripotent ◽  
Aavs1 Locus ◽  
High Level

Key Points When targeted to a single allele of the AAVS1 locus, the Gp1ba promoter drives a high level of expression specifically to megakaryocytes. Transgene rescue in iPSCs provides a model for the return of surface αIIbβ3 expression to near-normal levels in patients with type I GT.

Regulatory elements in the 5'-flanking region and the first intron contribute to transcriptional control of the mouse alpha 1 type I collagen gene

1989 ◽  
Vol 9 (5) ◽  
pp. 2224-2227
Author(s):  
R A Rippe ◽  
S I Lorenzen ◽  
D A Brenner ◽  
M Breindl
Keyword(s):  
Transcriptional Control ◽  
Type I Collagen ◽  
Regulatory Elements ◽  
Regulatory Sequences ◽  
Type I ◽  
Col1a1 Gene ◽  
First Intron ◽  
Flanking Region ◽  
Specific Regulation ◽  
High Level

We have identified two blocks of regulatory sequences located in the 5'-flanking region and the first intron of the mouse alpha 1 type I collagen (COL1A1) gene. Both blocks were found to contain positive as well as negative regulatory elements. Sequences located within 222 base pairs upstream of the transcription start site showed a strong stimulatory effect on the COL1A1 promoter and were sufficient for tissue-specific regulation of the COL1A1 gene. The combined upstream and intron regulatory sequences showed a marked inhibition of COL1A1 promoter activity in fibroblasts. This finding suggests that additional, more remote regulatory sequences may be required for establishing the high level of activity of the endogenous COL1A1 gene in fibroblastoid cells.

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