• Title/Summary/Keyword: sonication efficiency

Search Result 63, Processing Time 0.027 seconds

Optimization of different factors for an Agrobacterium-mediated genetic transformation system using embryo axis explants of chickpea (Cicer arietinum L.)

  • Sadhu, Suman Kalyan;Jogam, Phanikanth;Gande, Kranthikumar;Banoth, Raghu;Penna, Suprasanna;Peddaboina, Venkataiah
    • Journal of Plant Biotechnology
    • /
    • v.49 no.1
    • /
    • pp.61-73
    • /
    • 2022
  • In this study, we developed a reliable and efficient Agrobacterium-mediated genetic transformation system by applying sonication and vacuum infiltration to six chickpea cultivars (ICCV2, ICCV10, ICCV92944, ICCV37, JAKI9218, and JG11) using embryo axis explants. Wounded explants were precultured for 3 days in shoot induction medium (SIM) before sonication and vacuum infiltration with an Agrobacterium suspension and co-cultivated for 3 days in co-cultivation medium containing 100 µM/l of acetosyringone and 200 mg/l of L-cysteine. Responsive explants with putatively transformed shoots were selected using a gradual increase in kanamycin from 25 mg/l to 100 mg/l in selection medium to eliminate escapes. Results showed optimal transformation efficiency at a bacterial density of 1.0, an optical density at 600 nm wavelength (OD600), and an infection duration of 30 min. The presence and stable integration of the β-glucuronidase (gusA) gene into the chickpea genome were confirmed using GUS histochemical assay and polymerase chain reaction. A high transformation efficiency was achieved among the different factors tested using embryo axis explants of cv. JAKI 9218. Of the six chickpea cultivars tested, JAKI9218 showed the highest transformation efficiency of 8.6%, followed by JG11 (7.2%), ICCV92944 (6.8%), ICCV37 (5.4%), ICCV2 (4.8%), and ICCV10 (4.6%). These findings showed that the Agrobacterium-mediated genetic transformation system will help transfer novel candidate genes into chickpea.

Expression of Modified Green Fluorescent Protein in Suspension Culture of Taxus cuspidata

  • Kim, Chang-Heon;Kim, Kyung-Il;Chung, In-Sik
    • Journal of Microbiology and Biotechnology
    • /
    • v.10 no.1
    • /
    • pp.91-94
    • /
    • 2000
  • The suspension cells of Taxus cuspidata were transformed with Agrobacterium tumefaciens harboring binary vector pCAMBIE1302 encoding mgfp. Transient transfection efficiency was compared by using the fluoremetric measurement. The transient transfection efficiency was improved by transformation with DMSO and/or sonication treatment. Optimum conditions for DMSO and sonication treatment were 3% and 30sec, respectively. selection and maintenance of transformed cells were continued for 3 months. An insertion of the mgfp gene in transformed cells was detected by PCR and an expression of GFP confirmed by the western blot analysis.

  • PDF

A Comparison of Sonication and Microwave-assisted Extraction Method for Speciation of Arsenic in Fish Tissue, DORM-2 (어류중 비소의 종분화 분석을 위한 초음파 추출법과 마이크로파 추출법의 비교)

  • Yoon, Cheol-Ho;Park, Yong-Chul;Hong, Jong-Ki
    • Analytical Science and Technology
    • /
    • v.16 no.2
    • /
    • pp.134-142
    • /
    • 2003
  • Comparison of a microwave-assisted extraction with sonication extraction was performed for arsenic speciation in fish tissue with chromatographic separation and inductively coupled plasma mass spectrometry detection. The detection limits of arsenicals with ultrasonic nebulizerand cross-flow nebulizer were shown to be similar. The arsenicals investigated were arsenobetaine (AsB), arsenite [As(III)], dimethylarsine acid (DMA), monomethylarsonic acid (MMA), arsenate [As(v)], and phenylarsonic acid (PAA). Quantitative extraction of arsenicals from dogfish muscle, DORM-2, standard reference material of NRCC (National Research Council of Canada) was achieved using 50% (v/v) methanol-water in both extraction methods. Extraction efficiency of arsenobetaine in both methods is greater than 82% with RSDs on replicates of less than 5%. The concentrations of AsB determined in extract of microwave assisted extraction and sonication methods were $14.18{\pm}0.42mg\;kg^{-1}$ and $13.54 {\pm}0.84mg\;kg^{-1}$, respectively. And the concentrations of DMA were $0.45{\pm}0.06mg\;kg^{-1}$ and $0.44{\pm}0.06mg\;kg^{-1}$, respectively.

Manipulation of Surface Carboxyl Content on TEMPO-Oxidized Cellulose Fibrils

  • Masruchin, Nanang;Park, Byung-Dae
    • Journal of the Korean Wood Science and Technology
    • /
    • v.43 no.5
    • /
    • pp.613-627
    • /
    • 2015
  • Simple methods of conductometric titration and infrared spectroscopy were used to quantify the surface carboxyl content of cellulose fibrils isolated by 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-mediated oxidation. The effects of different cellulose sources, post or assisted-sonication oxidation treatment, and the amount of sodium hypochlorite addition on the carboxyl content of cellulose were reported. This study showed that post sonication treatment had no influence on the improvement of surface carboxyl charge of cellulose macrofibrils (CMFs). However, the carboxyl content increased for the isolated cellulose nanofibrils (CNFs). Thus the carboxyl content of CNFs is different from those of their corresponding bulk oxidized cellulose and CMFs. Filter paper as a CNF source imparted a higher surface charge than did hardwood bleached kraft pulp (HWBKP) and microcrystalline cellulose (MCC). It was considered that the crystallinity and microstructure of the initial cellulose affected oxidation efficiency. In addition, the carboxyl content of cellulose was successfully controlled by applying sonication treatment during the oxidation reaction and adjusting the amount of sodium hypochlorite.

Entrapment of Plasmid DNA in Liposomes (리포솜을 이용한 플라스미드 DNA의 봉입)

  • Song, Mi-Hyang;Lee, Mann-Hyung;Yong, Chul-Soon;Oh, Doo-Man
    • Journal of Pharmaceutical Investigation
    • /
    • v.26 no.4
    • /
    • pp.291-297
    • /
    • 1996
  • Liposomes of $pSV-{\beta}-Galactosidase$ vector plasmid DNA with various lipid composition were prepared by the thin-film method. Size distribution, shape and the efficiency of plasmid DNA encapsulation were investigated. Effect of sonication time on the plasmid DNA entrapment in liposomes and stability at $4^{\circ}C$ were also examined. Sizes of neutral liposomes were about 100-200 nm and above $1\;{mu}m$, and those of cationic liposomes were about 400-600 nm and above $1\;{mu}m$. Shapes of liposomes entrapped plasmid DNA were spherical. Proper sonication time for better entrapment was below 15 minutes and stability at $4^{\circ}C$ was decreased rapidly after 1 day. Plasmid DNA entrapments of complex liposomes of various lipids were higher than those of liposomes made from one sort of lipid. Plasmid DNA entrapments of cationic liposomes were higher than those of neutral liposomes.

  • PDF

The Effect of Reaction Temperature and Volume in the Sonolysis of 1,4-Dioxane (1,4-Dioxane의 초음파 처리시 반응 온도와 부칙의 영향)

  • Son, Hyun-Seok;Choi, Seok-Bong;Eakalak, Khan;Zoh, Kyung-Duk
    • Journal of Korean Society of Environmental Engineers
    • /
    • v.27 no.10
    • /
    • pp.1114-1122
    • /
    • 2005
  • This research investigates the effects of sonication mode, reaction temperature and volume on sonolysis of 1,4-dioxane in order to increase the degradation efficiency and kinetics. The degradation efficiency in case with pulse mode was about 10 % higher than that in case with continuous mode. The degradation profiles in both cases, which were performed without the control of reaction temperature and in 1000 mL, were composed of three steps. However, 1,4-D was mainly degraded in the initiation step as the first portion and the acceleration step as the second portion. The initial step agreed with zero-order expression well, while the acceleration step could be fitted with pseudo 1st-order expression. The kinetic model in case with $5^{\circ}C$ and 300 mL conformed to pseudo 1st-order, while that in cases with $10^{\circ}C$ to $40^{\circ}C$ agreed with zero-order expression. The degradation efficiency and profile of 1,4-D in the experiment with $20^{\circ}C$ and 300 mL was higher and simpler than that in case with $20^{\circ}C$ and 1000 mL. The reaction temperature and volume influence bubble intensity, which was produced in sonication. The increase of bubble intensity induced to augment the production of OH radical in sonication.

Feasibility Study on Remediation for Railroad-contaminated Soil with Waste-lubricant (윤활유 유래 철도 오염토양의 정화방법 연구)

  • Park, Sung-Woo;Shin, Min-Chul;Jeon, Chil-Sung;Baek, Ki-Tae;Lee, Jae-Young
    • Journal of Soil and Groundwater Environment
    • /
    • v.13 no.2
    • /
    • pp.30-35
    • /
    • 2008
  • In this study, the feasibility of soil washing, chemical oxidation and sonication was investigated to treat lubricantcontaminated railroad soil. Tergitol, a non-ionic surfactant, was used as a washing agent with or without iso-propyl acohol as a cosolvent. However, it was not effective to remove lubricant from soil even though tergitol was the most effective washing agent for diesel-contaminated soil. The cosolvent reduced the overall washing efficiency. Chemical oxidation removed 30% of lubricant from contaminated soil. Soil washing after chemical oxidation extracted additionally 16-17% of lubricant. Sonication enhanced-soil washing showed enhanced overall efficiency of soil washing. Lubricant-contaminated soil should be remediated by the other technology used for diesel-contaminated soil.

Effects of ultrasonication on the analysis of sulforaphane content in vegetables (초음파 처리가 채소 중의 sulforaphane 분석에 미치는 영향)

  • Choi, A-Reum;Lee, Gun-Soon;Chae, Hee-Jeong
    • Journal of the Korea Academia-Industrial cooperation Society
    • /
    • v.9 no.3
    • /
    • pp.794-799
    • /
    • 2008
  • The effect of ultrasonication on the sulforaphane analysis in cruciferous vegetables was investigated by GC/MS. The ultrasonication of the analysis samples was carried out in dichloromethane as a solvent, which was followed by concentration in nitrogen gas, and the analysis of sulforaphane was performed using selective ion monitoring (SIM) at m/z 72, 160, 55 and 114. The content of sulforaphane was the highest in the extract of broccoli (149 ppm), and followed by cabbage (67.9 ppm) and radish (35.4 ppm). When the vegetable samples were shaked after ultrasonication, the extraction efficiency of sulforaphane was 2.7-fold enhanced, compared to the extraction by shaking. The result suggested that pretreatment including sonication can be used fer improving the extraction efficiency of sulforaphane.

Cationic Emulsions with Galactosylated Chitosan as a Novel Gene Delivery System

  • Choi, Woo-Jeong;Choi, Sung-Hee;Ahn, Woong-Shick;Kim, Chong-Kook
    • Proceedings of the PSK Conference
    • /
    • 2002.10a
    • /
    • pp.427.1-427.1
    • /
    • 2002
  • To improve stability and transfection efficiency, a novel combination of cationic emulsion and galactosylated chitosan was developed for targeted gene delivery. Six formulations of cationic liposome and our novel emulsion were prepared for comparison of stability and transfection efficiency. Cationic liposomes composed of 3[N-(N.N dimethylaminoethylene) carbamoyl] cholesterol (DC-Chol) and dioleyl phophatidyl ethanolamine (DOPE) were prepared by extrusion method and cationic emulsions composed of DC-Chol. DOPE. castor oil, and Tween 80 were prepared by sonication method. (omitted)

  • PDF

A Study on the Factors Affecting Entrapment Efficiency and Particle Size of Ethosomes (Ethosomes의 포집효율과 입자크기에 영향을 주는 인자에 관한 연구)

  • Jin, Byung Suk;Lee, Sang Mook;Lee, Kwang Hee
    • Applied Chemistry for Engineering
    • /
    • v.17 no.2
    • /
    • pp.138-143
    • /
    • 2006
  • Ethosome is a liquid crystalline vesicle prepared by hydration of ethanol-dissolved lecithin with a solution containing hydrophilic components. Investigation of factors affecting the entrapment efficiency and particle size of ethosomes was carried out, because the high entrapment efficiency and small particle size are prerequisite in developing ethosomes as a drug delivery system. The variations of properties of ethosomes with constituent composition and preparation method were examined using a calcein as a hydrophilic marker. It was observed that the amount of ethanol and calcein solution, phosphatidyl choline content in lecithin, preparation temperature, stirring rate, and PBS addition method had a considerable effect on the properties of ethosome. Sonication treatment resulted in the reduction of entrapment efficiency of ethosome, which was due to the release of entrapped components in the vesicles by strong sonication vibration.