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Development of Industrial Strain, Medium Characteristics and Biochemical Pathways
Published in Debabrata Das, Soumya Pandit, Industrial Biotechnology, 2021
Protoplast fusion: This is a physical phenomenon which can be used for both prokaryotic and eukaryotic cells. Most of the filamentous fungi are made industrially viable through protoplasmic fusion. Protoplast refers to whole cells except for the cell wall. The cell wall of the bacteria is cleaved by lytic agents such as cellulase, lysozymes or macerozyme and with the presence of fusion agents two or more protoplasts having physical contact fuse together. The surface potential of the membrane is altered when it is brought into close proximity which ultimately results in fusion. Through this process interspecies genes can also be fused together to get the desirable properties of the corresponding strain. Transformation efficiency is also more than 80% (Chen et al., 2010).
Basic Molecular Cloning of DNA and RNA
Published in Jay L. Nadeau, Introduction to Experimental Biophysics, 2017
The fraction of the transformed competent cells that should be plated to get a good number of colonies, not a lawn, can be estimated by adjusting the amount of DNA used and using the published transformation efficiency (T) of the strain. This is defined as the number of colonies obtained per microgram of DNA. Typical DNA concentrations used per reaction are 1–10 ng; more than 10 ng usually does not result in more colonies. Thus, the number of colonies obtained (N) is given by
Establishment and elicitation of transgenic root culture of Plantago lanceolata and evaluation of its anti-bacterial and cytotoxicity activity
Published in Preparative Biochemistry & Biotechnology, 2021
Samaneh Rahamouz-Haghighi, Khadijeh Bagheri, Ali Sharafi, Hossein Danafar
The transformation efficiency of many plants like Withania somnifera[50] and Rauwolfia serpentine[51] has increased in presence of Acetosyringone. Muthusamy et al. employed 75 µM of Acetosyringone and indicated that the presence of Acetosyringone increased the efficiency of transformation up to 90% while in the absence of Acetosyringone the transformation efficiency was 36.6%.[52] In another research, Bhagat et al. stated that applying 125 µM of Acetosyringone, the transformation efficiency increased up to 55% whereas in the absence of Acetosyringone, the transformation efficiency was around 20%.[51] In the same line, Amoah and colleagues obtained an increased number of explants producing blue spots with 200 µM Acetosyringone.[53] Paul et al. used different concentrations of Acetosyringone to evaluate optimal transformation efficiency. According to their results, 150 µM concentration determined as an optimal condition[54] However, they pointed out that Acetosyringone at a concentration up to 200 µM was considered to be nontoxic to Agrobacterium, but reduced transient transformation efficiency suggesting 150 µM was optimal for patchouli leaf transformation.[55]
Effects of carbon source, C/N ratio, nitrate, temperature, and pH on N2O emission and functional denitrifying genes during heterotrophic denitrification
Published in Journal of Environmental Science and Health, Part A, 2018
Yun-Yeong Lee, Hyungjoo Choi, Kyung-Suk Cho
The effect of the C/N ratio on transformation efficiency is shown in Figure 3(a). At a C/N ratio of 0, the removal efficiency of NO3−-N was 5.8 ± 0.00% with most NO3−-N (94.2 ± 0.00%) remaining in the system. The removed NO3−-N (5.8 ± 0.00%) was reduced to N2O-N (0.8 ± 0.55%) and N2-N (5.0 ± 0.67%). At a C/N ratio of 1.28, 73.9 ± 4.11% of NO3−-N was reduced to 28.2 ± 0.29% N2O-N and 45.7 ± 7.71% N2-N. When the C/N ratio was 2.57, all removed NO3−-N (53.1 ± 3.04%) was reduced to N2O-N (53.1 ± 3.04%) and N2 gas was not generated. At C/N ratios of 5.14 and 12.85, the removal efficiency of NO3−-N was 100% and it was completely reduced to N2 gas, which is the final product of the denitrification process.
Genetic and substrate-level modulation of Bacillus subtilis physiology for enhanced extracellular human interferon gamma production
Published in Preparative Biochemistry and Biotechnology, 2018
Nitin Kumar, Rajat Pandey, Ashish Anand Prabhu, Veeranki Venkata Dasu
In B. subtilis, the natural competency develops under nutrient depletion condition.[42] Such stage predominantly arises just after the end of the exponential phase and before the commencement of stationary phase. It is also marked by instantaneous and distinct intensification in cell motility. To capture the cells at this stage, the active overnight culture of B. subtilis WB800N was inoculated in medium A and growth profile was observed along with microscopy after every 20 min (supplementary Figure S2(b)). The culture picked up motility after 200 min at 1.45 O.D.650nm and the motility was intact for the next 1 hr with the peak motility at 260 min with 2.1 O.D.650nm. Within this 1 hr, 50 µL of culture was taken at three different time points 220, 240, and 260 min and inoculated in 450 µL of fresh medium A supplemented with CaCl2 and MgSO4. The culture in the fresh medium attained a higher density biomass and higher motility after a further incubation of 80–90 min. At this point, the highly motile and competent culture was transformed with 10 ng of pIFNG, pcoIFNG, and pcoIFNGhis an transformation efficiency of 70 (±4) cfu/10 ng of the plasmid was observed for all the three plasmids. The positive clones were screened by plating on LB with Neomycin (10 µg mL−1) and chloramphenicol (10 µg mL−1) and the strains were designated as BSIFNG, BScoIFNG, and BScoIFNGhis correspondingly. The 260-min culture resulted in highest transformation efficiency.