scholarly journals OPTIMIZATION OF THE CULTURE MEDIUM COMPOSITION TO INCREASE THE BIOSYNTHESIS OF RECOMBINANT HUMAN INTERLEUKIN-7 IN ESCHERICHIA COLI

2020 ◽  
Vol 9 (4) ◽  
pp. 761-768
Author(s):  
Valentyna Motronenko
Keyword(s):  
Escherichia Coli ◽  
Culture Medium ◽  
Medium Composition ◽  
Interleukin 7

scholarly journals The Effects of Temperature and Culture Medium Composition on Protein Recombinant JSU-pET Expression

2019 ◽  
Vol 17 (2) ◽  
pp. 31
Author(s):  
Indriawati Indriawati ◽  
Slamet Diah Volkandari ◽  
Endang Tri Margawati
Keyword(s):  
Escherichia Coli ◽  
Protein Expression ◽  
Culture Medium ◽  
Medium Composition ◽  
Protein Yield ◽  
Culture Temperature ◽  
Factors Influencing ◽  
Effects Of Temperature ◽  
The Right ◽  
Medium Culture

There are several factors influencing the protein expression. The study was aimed to increase the yield of protein recombinant JSU-pET by applying combination of temperature and medium culture composition. This research was composed in a 2x2 factorial design with two level temperatures of 37<sup>o</sup>C and 25<sup>o</sup>C and 2 medium compositions of LB and mLB (modified). <em>Escherichia coli</em> bearing JSU-pET plasmid were grown in two applied media and incubated in two applied temperatures. The cells were harvested by centrifugation the pellets were then lysed. The lysed cells were centrifuged, the supernatant then purified by Ni-NTA Resin Agarose. The purified JSU protein recombinant was characterized and quantified by spectrophotometer. The Result showed that the JSU protein recombinant was successfully expressed on the right size (37kDa) in all treatments even though the JSU-pET showed sharper band in m LB at 37<sup>o</sup>C. Statistically the protein yield was not significant different (P&gt;0.05), however culture temperature of 25<sup>o</sup>C showed higher yield (0.618±0.095 vs 0.704±0.094) than cultured at 37<sup>o</sup>C (0.598±0.137 vs 0.553±0.041), respectively either cultured in LB or mLB. This finding suggests that a lower culture temperature could increase protein yield both either in LB or modified LB.

scholarly journals Boosting Auto-induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives

2021 ◽  
Vol 28 ◽  
Author(s):  
Nariyasu Tahara ◽  
Itaru Tachibana ◽  
Kazuyo Takeo ◽  
Shinji Yamashita ◽  
Atsuhiro Shimada ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Proteins ◽  
Culture Medium ◽  
Thermus Thermophilus ◽  
Medium Composition ◽  
Final Concentration ◽  
Induction Medium ◽  
Lac Operon ◽  
Escherichia Coli Expression ◽  
Complex Culture

Background: Auto-induction is a convenient way to produce recombinant proteins without inducer addition using lac operon-controlled Escherichia coli expression systems. Auto-induction can occur unintentionally using a complex culture medium prepared by mixing culture substrates. The differences in culture substrates sometimes lead to variations in the induction level. Objectives: In this study, we investigated the feasibility of using glucose and lactose as boosters of auto-induction with a complex culture medium. Method: First, auto-induction levels were assessed by quantifying recombinant GFPuv expression under the control of the T7lac promoter. Effectiveness of the additive-containing medium was examined using ovine angiotensinogen (tac promoter-based expression) and Thermus thermophilus manganese-catalase (T7lac promoter-based expression). Results: Auto-induced GFPuv expression was observed with the enzymatic protein digest Polypepton, but not with another digest tryptone. Regardless of the type of protein digest, supplementing Terrific Broth medium with glucose (at a final concentration of 2.9 g/L) and lactose (at a final concentration of 7.6 g/L) was successful in obtaining an induction level similar to that achieved with a commercially available auto-induction medium. The two recombinant proteins were produced in milligram quantity of purified protein per liter of culture. Conclusion: The medium composition shown in this study would be practically useful for attaining reliable auto-induction for E. coli-based recombinant protein production.

Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Water Concentration ◽  
Medium Composition ◽  
Coconut Water ◽  
Orchid Species ◽  
Randomized Design ◽  
Wet Weight ◽  
Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

Differentiation of mesenchymal stem cells from humans and animals into insulin-producing cells: An overview in vitro induction forms

2020 ◽  
Vol 16 ◽  
Author(s):  
Bruna O. S. Câmara ◽  
Bruno M. Bertassoli ◽  
Natália M. Ocarino ◽  
Rogéria Serakides
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Culture Medium ◽  
Adipogenic Differentiation ◽  
Medium Composition ◽  
Cell Therapies ◽  
Insulin Producing Cells ◽  
Msc Differentiation

The use of stem cells in cell therapies has shown promising results in the treatment of several diseases, including diabetes mellitus, in both humans and animals. Mesenchymal stem cells (MSCs) can be isolated from various locations, including bone marrow, adipose tissues, synovia, muscles, dental pulp, umbilical cords, and the placenta. In vitro, by manipulating the composition of the culture medium or transfection, MSCs can differentiate into several cell lineages, including insulin-producing cells (IPCs). Unlike osteogenic, chondrogenic, and adipogenic differentiation, for which the culture medium and time are similar between studies, studies involving the induction of MSC differentiation in IPCs differ greatly. This divergence is usually evident in relation to the differentiation technique used, the composition of the culture medium, the cultivation time, which can vary from a few hours to several months, and the number of steps to complete differentiation. However, although there is no “gold standard” differentiation medium composition, most prominent studies mention the use of nicotinamide, exedin-4, ß-mercaptoethanol, fibroblast growth factor b (FGFb), and glucose in the culture medium to promote the differentiation of MSCs into IPCs. Therefore, the purpose of this review is to investigate the stages of MSC differentiation into IPCs both in vivo and in vitro, as well as address differentiation techniques and molecular actions and mechanisms by which some substances, such as nicotinamide, exedin-4, ßmercaptoethanol, FGFb, and glucose, participate in the differentiation process.

Study on Optimization of Fermentation Condition for Nitrilase-Producer Escherichia Coli BL21 (DE3)/pET-Nit

2011 ◽  
Vol 175-176 ◽  
pp. 192-196 ◽  
Author(s):  
Li Li Feng ◽  
Jian Fei Zhang ◽  
Hui Luo ◽  
Zheng Li ◽  
Hong Jie Zhang
Keyword(s):  
Escherichia Coli ◽  
Recombinant Strain ◽  
Culture Medium ◽  
Culture Conditions ◽  
Fermentation Condition ◽  
Hydrogen Phosphate ◽  
Strain Bl21 ◽  
Initial Ph ◽  
Potassium Hydrogen ◽  
Optimization Of Fermentation

The paper concentrated on the optimization of the recombinant strain BL21 (DE3)-PE7-Nit. The component of culture medium and the culture conditions were optimized. The optimized medium was: yeast extract 10 g/l, L-glutamate sodium 8 g/l, MgSO4.7H2O 0.7 g/l, Isopropyl-β-D-thiogalactopyranoside 0.3 mmol/L, potassium hydrogen phosphate 0.5 g / L, phosphate Potassium 0.5 g / L and the culture condition was: initial pH 7.0, inoculum 2%. The result showed that the activity of nitrilase prepared with these conditions increased by 130.37 % through optimization.

P–218 Analysis of the occurrence of microbial contamination in IVF culture system and the effect of microorganisms on embryo development and clinical outcomes

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
F Du ◽  
R Li ◽  
Q Zhang ◽  
W Wang
Keyword(s):  
Escherichia Coli ◽  
In Vitro Culture ◽  
Clinical Outcomes ◽  
Embryo Transfer ◽  
Follicular Fluid ◽  
Culture System ◽  
Microbial Contamination ◽  
Culture Medium ◽  
Ivf Et

Abstract Study question what is the source, prevalence, and influence of microbial contamination on in vitro fertilization (IVF) and embryo transfer (ET) cycles? Summary answer Microbial contamination mainly occurs on Day 2, most caused by Escherichia coli carried with semen. ICSI could prevent contamination effectively and get good clinical outcomes. What is known already Microbial contamination occurs in IVF-ET system occasionally, which is hard to stop happening. The IVF culture system and laboratory environment, the patients’ follicular fluid and semen are not absolutely sterile, while the antibiotics in culture medium isn’t effective for all microbe types, and the artificial operations may bring in microbes. Generally, microbial contamination leads to degradation of embryos, reduction the number of embryos available, and infection of female reproductive tract, which would increase the cost of patients’ time, money, and bring psychological damages. A better understanding of embryo contamination in IVF culture system is of added value. Study design, size, duration A total of 29583 IVF-ET cycles were enrolled in this prospective observational study, from January 2010 to December 2020, included 70 microbial contamination cycles discovered in Day1-Day3 (D1-D3) of in vitro culture. Follicular fluid and semen saved on oocyte retrieval day, and culture medium contaminated were examined and identified for microorganisms at each contamination cycle. Participants/materials, setting, methods Compared the contamination rate of different insemination methods (IVF/ICSI/IVF+ICSI), different in vitro culture days (D1-D3), and different samples examination (follicular fluid, semen, culture medium) respectively, identified the source of microorganism types, compared the IVF culture outcomes and clinical outcomes between total contamination group (TC group, 42 cases) and partial contamination group (PC group, 28 cases). Main results and the role of chance A total of 70 microbial contamination cases occurred in 29583 oocyte retrieving cycles (0.24%), and it was observed only in IVF embryos but never in ICSI (Intracytoplasmic sperm injection) embryos. 38 contamination cases occurred on D2 with a highest ratio (54.3%) compared to D1 (32.9%) and D3(12.9%); Compared with follicular fluid, semen was the main cause inducing contamination from D1 to D3, and Escherichia coli in semen and culture medium, Enterococcus faecalis in follicular fluid proved to be the most common sources. Compared with TC group, the PC group showed a lower rate of No-available embryos (21.4% vs 81.0%) and a higher rate of blastocyst formation (41.2% vs 28.6%), In addition, the clinical pregnancy rate of PC group was higher than that of TC group in both fresh and frozen-thawed embryo transfer cycles (31.3% vs 16.7%, 38.5% vs 0.0%). Limitations, reasons for caution Further study is still necessary to better understand the sources that induce microbial contamination embryos, and more efficient methods are required to remove the microbes on these contaminated embryos so as better develop and manage a sterile micro-environment for successful embryo growth. Wider implications of the findings: The differential embryonic microbe types associated to different IVF culture and clinical outcomes in patients undergoing IVF-ET might have profound implications for understanding the microbial sources and making a better management of IVF culture system. Trial registration number Not applicable

scholarly journals Efficiency of using different types induction culture medium for hexaploid triticale anther cultivation

2019 ◽  
Vol 25 ◽  
pp. 260-265
Author(s):  
E. V. Lagunovskaya ◽  
O. I. Zaitseva ◽  
V. A. Lemesh
Keyword(s):  
Culture Medium ◽  
Medium Composition ◽  
Induction Medium ◽  
Hexaploid Triticale ◽  
Culture Methods ◽  
Study Results ◽  
Short Period ◽  
Androgenesis In Vitro ◽  
The Republic

Aim. Triticale is one of the main grain crops of the Republic of Belarus. Further progress in the selection of this culture involves the accelerated creation of highly productive early ripening varieties resistant to abiotic and biotic factors. The method of induced androgenesis in vitro makes it possible to obtain stable homozygous lines in a short period of time and to eliminate the lengthy process of inbreeding used in classical breeding to fix the desired traits. Methods. The tissue and cell culture methods for plants was used in the study. Results. The influence of the induction medium composition on the efficiency of in vitro induced androgenesis in varieties and lines of hexaploid triticale is assessed. The influence of three types of induction culture medium, the type of phytohormones and the presence or absence of cefotaxime in the medium are analyzed. Results. It has been shown that using the C-17 culture medium supplemented with 2.0 mg/l 2,4-D and 0.5 mg/l kinetin without adding cefotaxime is most effective for the anther triticale cultivation. Keywords: triticale, anther culture, induction nutrient medium, embryoids, calli, regenerant plants, cefotaxime.

scholarly journals The Isolation and Oharacterization of An Rna Bacteriophage

1964 ◽  
Vol 17 (3) ◽  
pp. 719 ◽  
Author(s):  
CI Davern
Keyword(s):  
Escherichia Coli ◽  
Dna Synthesis ◽  
Culture Medium ◽  
Buoyant Density ◽  
Sedimentation Coefficient ◽  
Caesium Chloride ◽  
Specific Phage ◽  
K 12 ◽  
Male Specific ◽  
Rna Phage

An enrichment procedure for the isolation of RNA bacteriophage is described. The method involves the inoculation of sewage samples into cultures of Escherichia coli K-12 Hfr under conditions where DNA synthesis is restricted by the addition of 5-fiuorodeoxyuridine to the culture medium. Six phage isolates were made and all of them were shown to be male-specific. One of the male-specific phage was further characterized as an RNA phage, having very similar properties to RNA phage already isolated in other parts oftha world. This RNA phage has a buoyant density of 1�42 g/cm3 in caesium chloride. and has a sedimentation coefficient of 79'5 Sin O'Ol:M Tria-HOI buffer, pH 7� 4, at 20�0.

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