scholarly journals A concise study on the expression of GFP protein in E. coli strain DH5α

2021 ◽  
Vol 16 (3) ◽  
Author(s):  
Maryada Garg ◽  
Anoop K. Dobriyal
Keyword(s):  
Protein Gene ◽  
Bacterial Culture ◽  
Research Work ◽  
Coli Strain ◽  
Foreign Gene ◽  
E Coli ◽  
Biotechnology Research ◽  
Indispensable Tool ◽  
The Given ◽  
Standard Grade

The GFP protein is a protein of high interest for molecular biologists and biotechnologists. Since 1994, this protein has proved to be an indispensable tool for molecular biology and biotechnology research work. This protein requires only oxygen and an energy source like glucose to work. It gives a green colour in presence of UV to blue light. This protein can be attached with the foreign gene to track its expression. The main aim of the task is to introduce GFP into the given bacterial culture. Induction of the protein gene is followed by Bradford assay; quantification of protein is done using this. The results of gene induction are checked via SDSPAGE and western blot. All preparations are of standard grade and all readings are taken in triplicates. A standard graph is also made to find out the protein in the unknown.

scholarly journals Cation Transport in Escherichia coli

1961 ◽  
Vol 45 (2) ◽  
pp. 355-369 ◽  
Author(s):  
Stanley G. Schultz ◽  
A. K. Solomon
Keyword(s):  
Growth Medium ◽  
Bacterial Culture ◽  
Coli Strain ◽  
Metabolic Energy ◽  
Logarithmic Phase ◽  
Concentration Gradients ◽  
E Coli ◽  
Ion Movements ◽  
K Concentration ◽  
K 12

Methods have been developed to study the intracellular Na and K concentrations in E. coli, strain K-12. These intracellular cation concentrations have been shown to be functions of the extracellular cation concentrations and the age of the bacterial culture. During the early logarithmic phase of growth, the intracellular K concentration greatly exceeds that of the external medium, whereas the intracellular Na concentration is lower than that of the growth medium. As the age of the culture increases, the intracellular K concentration falls and the intracellular Na concentration rises, changes which are related to the fall in the pH of the medium and to the accumulation of the products of bacterial metabolism. When stationary phase cells, which are rich in Na and poor in K, are resuspended in fresh growth medium, there is a rapid reaccumulation of K and extrusion of Na. These processes represent oppositely directed net ion movements against concentration gradients, and have been shown to be dependent upon the presence of an intact metabolic energy supply.

Deadly E. Coli Strain May Evade EPA's Test

2011 ◽  
pp. 020811151303
Author(s):  
Sara Peach
Keyword(s):  
Coli Strain ◽  
E Coli

Transport of escherichia coli through soil to groundwater traced by randomly amplified polymorphic DNA (RAPD)

1997 ◽  
Vol 35 (11-12) ◽  
pp. 351-357 ◽  
Author(s):  
R. Rothmaier ◽  
A. Weidenmann ◽  
K. Botzenhart
Keyword(s):  
Negative Impact ◽  
Random Amplified Polymorphic Dna ◽  
Coli Strain ◽  
Soil Samples ◽  
Test Field ◽  
Liquid Manure ◽  
E Coli ◽  
Rapd Pattern ◽  
Geological Situation ◽  
Different Strains

Isolates (50) of E. coli obtained from liquid manure (20 bovine, 20 porcine) were genotyped using random amplified polymorphic DNA (RAPD). Typing revealed 9 and 14 different strains in bovine and porcine liquid manure respectively with no strains in common. One porcine strain, showing a simple RAPD pattern, was subcultured and spread on a test field (1.5l/m2 at 1010 cfu/l) in a drinking water protection zone with loamy to sandy sediments in the Donauried area, Baden-Wurttemberg. Soil samples and groundwaters were collected at monthly intervals October 1994 – June 1995 during which 114 E. coli isolates were recovered. The first occurrence and maximum concentration of E. coli in soil samples taken from more than 20cm depth was in January 1995, declining rapidly with depth and time. All isolates from soil and only one from groundwater showed the RAPD pattern of the spread E. coli strain. The results could not demonstrate a severe negative impact of the spreading of liquid manure on the bacteriological quality of the groundwater in the given geological situation. The distinct strain patterns found in different kinds of liquid manure suggest that genotyping of E. coli by RAPD may be an adequate tool for tracing sources of faecal contamination.

Berberine Chloride Dihydrate Enthused Nanovesicles for the Management of Dermatitis

2020 ◽  
Vol 10 ◽  
Author(s):  
Nimisha Srivastava ◽  
Zeeshan Fatima ◽  
Chanchal Deep Kaur ◽  
Dilshad Ali Rizvi
Keyword(s):  
Laser Scanning ◽  
Skin Irritation ◽  
Research Work ◽  
Slight Modification ◽  
Entrapment Efficiency ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Chloride Dihydrate ◽  
E Coli ◽  
Berberine Chloride

Background: Dermatitis is a common inflammatory skin disease that is affecting up to 25% of children and 1%-3% of adults worldwide. Paucity of exact cure for dermatitis and untoward side effects of topical immunosuppressive steroids has resulted into a great need for making use of complementary medicine to treat dermatitis. Objective: The present research work involved the development of Berberine chloride dihydrate (BCD) enthused nanovesicles i.e. ethosomes for the management of dermatitis. Method: Ethosomes were prepared by slight modification of cold method using varying concentrations of SPC (1-3%) and ethanol (10-40%) Optimized batch BCD 12 was further added to Carbopol 934P for gel formation. GEL BCD 12 was subjected to “anti-bacterial, dermatitis and skin irritation study. Result: The vesicles were in size range 142.42-398.31 nm while polydispersity index (PDI) ranges from 0.114-1.56 and for zeta potential it was from-18.8 to -39.4. Entrapment efficiency was from 46.05-88.79 %. Confocal laser scanning microscopy showed penetration depth of rhodamine enthused ethosome across rat skin upto 110 µm which was significantly higher than rhodamine solution (10 µm). In the anti-bacterial study, BCD loaded ethosomal gel (EG) showed maximum zone of inhibition of 18.5 mm against E. coli, 14.5 mm against P. aeruginosa and 23.0 mm against S. aureus. In dinitrochlorobenzene (DNCB) induced mice dermatitis model histopathology study showed marked decrease in amount of inflammatory cell nucleus in mice treated with BCD loaded ethosomal gel followed by 56% and 50 % increase in ear swelling and ear mass respectively in morphology study. Conventional marketed formulation showed nominal decrease in epidermal thickness, 66.67 % increase in ear thickness and 63.64 % increase in ear mass. Further Primary irritation index was less than 0.4 indicating negligible irritation in all the groups. Conclusion: It can be concluded that ethosomal gel is not only an efficient carrier for BCD but also proves its potential for the management of dermatitis.

scholarly journals Low endotoxin E. coli strain-derived plasmids reduced rAAV vector-mediated immune responses both in vitro and in vivo.

2021 ◽  
Author(s):  
Qingyun Zheng ◽  
Tianyi Wang ◽  
Xiangying Zhu ◽  
Xiao Tian ◽  
Chen Zhong ◽  
...  
Keyword(s):  
Immune Responses ◽  
Coli Strain ◽  
Raav Vector ◽  
E Coli

scholarly journals Regulation of Expression of the TIR-Containing Protein C Gene of the Uropathogenic Escherichia coli Strain CFT073

Pathogens ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 549
Author(s):  
Julia Ittensohn ◽  
Jacqueline Hemberger ◽  
Hannah Griffiths ◽  
Maren Keller ◽  
Simone Albrecht ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein C ◽  
Interleukin 1 ◽  
Coli Strain ◽  
Uropathogenic Escherichia Coli ◽  
Reporter Construct ◽  
Regulation Of Expression ◽  
E Coli ◽  
Strain Cft073 ◽  
Myeloid Differentiation Factor

The uropathogenic Escherichia coli strain CFT073 causes kidney abscesses in mice Toll/interleukin-1 receptor domain-containing protein C (TcpC) dependently and the corresponding gene is present in around 40% of E. coli isolates of pyelonephritis patients. It impairs the Toll-like receptor (TLR) signaling chain and the NACHT leucin-rich repeat PYD protein 3 inflammasome (NLRP3) by binding to TLR4 and myeloid differentiation factor 88 as well as to NLRP3 and caspase-1, respectively. Overexpression of the tcpC gene stopped replication of CFT073. Overexpression of several tcpC-truncation constructs revealed a transmembrane region, while its TIR domain induced filamentous bacteria. Based on these observations, we hypothesized that tcpC expression is presumably tightly controlled. We tested two putative promoters designated P1 and P2 located at 5′ of the gene c2397 and 5′ of the tcpC gene (c2398), respectively, which may form an operon. High pH and increasing glucose concentrations stimulated a P2 reporter construct that was considerably stronger than a P1 reporter construct, while increasing FeSO4 concentrations suppressed their activity. Human urine activated P2, demonstrating that tcpC might be induced in the urinary tract of infected patients. We conclude that P2, consisting of a 240 bp region 5′ of the tcpC gene, represents the major regulator of tcpC expression.

Research work undertaken in the Plant Transgenic Design Initiative in the Gene Research Center within Tsukuba Plant Innovation Research Center

Impact ◽  
2020 ◽  
Vol 2020 (3) ◽  
pp. 6-8
Author(s):  
Kazuo Watanabe
Keyword(s):  
Climate Change ◽  
Research Work ◽  
Plant Biotechnology ◽  
Research Center ◽  
Plant Genetics ◽  
Environmental Challenges ◽  
Innovation Research ◽  
Biotechnology Research ◽  
Genetically Improved ◽  
The University

The burgeoning area of plant genetics may hold the key to overcoming some of the most pressing environmental challenges. For example, crops can be genetically improved to make them better able to adapt to climate change, while genetic engineering of crops could help to address food security challenges. As such, a comprehensive understanding of plant genetics may enable humankind to make headway in addressing climate change and resulting challenges. Research in this area is therefore paramount. Research work undertaken in the Plant Transgenic Design Initiative (PTraD) in the Gene Research Center (GRC) within Tsukuba Plant Innovation Research Center (T-PIRC), located at the University of Tsukuba in Japan, is focused on plant sciences and biotechnologies. The PTraD is the centre of excellence in plant biotechnology research in Japan, shedding light on plant genetics and how this can be harnessed to solve environmental challenges such as climate change.

scholarly journals Novel Bacterial Production of Two Different Bioactive Forms of Human Stem-Cell Factor

2021 ◽  
Vol 22 (12) ◽  
pp. 6361
Author(s):  
Eunyoung Lee ◽  
Michelle Novais de Paula ◽  
Sangki Baek ◽  
Huynh Kim Khanh Ta ◽  
Minh Tan Nguyen ◽  
...  
Keyword(s):  
Stem Cell ◽  
Ion Exchange ◽  
Stem Cell Factor ◽  
Transmembrane Domain ◽  
Coli Strain ◽  
Exchange Chromatography ◽  
Soluble Form ◽  
Human Stem Cell ◽  
E Coli ◽  
Human Stem Cell Factor

Human stem-cell factor (hSCF) stimulates the survival, proliferation, and differentiation of hematopoietic cells by binding to the c-Kit receptor. Various applications of hSCF require the efficient and reliable production of hSCF. hSCF exists in three forms: as two membrane-spanning proteins hSCF248 and hSCF229 and truncated soluble N-terminal protein hSCF164. hSCF164 is known to be insoluble when expressed in Escherichia coli cytoplasm, requiring a complex refolding procedure. The activity of hSCF248 has never been studied. Here, we investigated novel production methods for recombinant hSCF164 and hSCF248 without the refolding process. To increase the solubility of hSCF164, maltose-binding protein (MBP) and protein disulfide isomerase b’a’ domain (PDIb’a’) tags were attached to the N-terminus of hSCF164. These fusion proteins were overexpressed in soluble form in the Origami 2(DE3) E. coli strain. These solubilization effects were enhanced at a low temperature. His-hSCF248, the poly-His tagged form of hSCF248, was expressed in a highly soluble form without a solubilization tag protein, which was unexpected because His-hSCF248 contains a transmembrane domain. hSCF164 was purified using affinity and ion-exchange chromatography, and His-hSCF248 was purified by ion-exchange and gel filtration chromatography. The purified proteins stimulated the proliferation of TF-1 cells. Interestingly, the EC50 value of His-hSCF248 was 1 pg/mL, 100-fold lower than 9 ng/mL hSCF164. Additionally, His-hSCF248 decreased the doubling time, increased the proportion of S and G2/M stages in the cell cycle, and increased the c-Myc expression at a 1000-fold lower concentration than hSCF164. In conclusion, His-hSCF248 was expressed in a soluble form in E. coli and had stronger activity than hSCF164. The molecular chaperone, MBP, enabled the soluble overexpression of hSCF164.

Urban wastewater disinfection for agricultural reuse: effect of solar driven AOPs in the inactivation of a multidrug resistant E. coli strain

2015 ◽  
Vol 178 ◽  
pp. 65-73 ◽  
Author(s):  
Giovanna Ferro ◽  
Antonino Fiorentino ◽  
María Castro Alferez ◽  
M. Inmaculada Polo-López ◽  
Luigi Rizzo ◽  
...  
Keyword(s):  
Multidrug Resistant ◽  
Coli Strain ◽  
Urban Wastewater ◽  
E Coli ◽  
Wastewater Disinfection
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