• Journal of Life Science
• /
• v.22 no.9
• /
• pp.1152-1158
• /
• 2012

Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene ($D{\times}5$) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the $D{\times}5$ and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring $D{\times}5$ or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with $D{\times}5$ gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the $D{\times}5$ and HPTII genes into the rice genome. We reconfirmed integration of the $D{\times}5$ and HPTII genes into the rice genome by Southern blot analysis. Wheat $D{\times}5$ transcripts in $T_1$ rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the $D{\times}5$ gene were successfully screened at the $T_1$ generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.

• Plant Biotechnology Reports
• /
• v.5 no.2
• /
• pp.147-156
• /
• 2011

Efficient Agrobacterium-mediated genetic transformation of Scoparia dulcis L. was developed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301 with ${\beta}$-glucuronidase (GUS) (uidA) and hygromycin phosphotransferase (hpt) genes. Two-day precultured leaf segments of in vitro shoot culture were found to be suitable for cocultivation with the Agrobacterium strain, and acetosyringone was able to promote the transformation process. After selection on shoot organogenesis medium with appropriate concentrations of hygromycin and carbenicillin, adventitious shoots were developed on elongation medium by twice subculturing under the same selection scheme. The elongated hygromycin-resistant shoots were subsequently rooted on the MS medium supplemented with $1mg\;l^{-1}$ indole-3-butyric acid and $15mg\;l^{-1}$ hygromycin. Successful transformation was confirmed by PCR analysis using uidA- and hpt-specific primers and monitored by histochemical assay for ${\beta}$-GUS activity during shoot organogenesis. Integration of hpt gene into the genome of transgenic plants was also verified by Southern blot analysis. High transformation efficiency at a rate of 54.6% with an average of $3.9{\pm}0.39$ transgenic plantlets per explant was achieved in the present transformation system. It took only 2-3 months from seed germination to positive transformants transplanted to soil. Therefore, an efficient and fast genetic transformation system was developed for S. dulcis using an Agrobacterium-mediated approach and plant regeneration via shoot organogenesis, which provides a useful platform for future genetic engineering studies in this medicinally important plant.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.6
• /
• pp.574-581
• /
• 2011

A Gram-positive bacterium was isolated from the saline soils of Jangpura (U.P.), India, and showed high-level of radiation-resistant property and survived upto 12.5 kGy dose of gamma radiation. The 16S rDNA sequence of this strain was examined, identified as Bacillus sp. strain HKG 112, and was submitted to the NCBI GenBank (Accession No. GQ925432). The mechanism of radiation resistance and gene level expression were examined by proteomic analysis of whole-cell extract. Two proteins, 38 kDa and 86.5 kDa excised from SDS-PAGE, which showed more significant changes after radiation exposure, were identified by MALDI-TOF as being flagellin and S-layer protein, respectively. Twenty selected 2-DE protein spots from the crude extracts of Bacillus sp. HKG 112, excised from 2- DE, were identified by liquid chromatography mass spectrometry (LC-MS) out of which 16 spots showed significant changes after radiation exposure and might be responsible for the radiation resistance property. Our results suggest that the different responses of some genes under radiation for the expression of radiation-dependent proteins could contribute to a physiological advantage and would be a significant initial step towards a fullsystem understanding of the radiation stress protection mechanisms of bacteria in different environments.

• Asian-Australasian Journal of Animal Sciences
• /
• v.33 no.5
• /
• pp.712-721
• /
• 2020

Objective: The Wannan Black pig is a typical Chinese indigenous, disease-resistant pig breed with high fertility, and a crude-feed tolerance that has been bred by artificial selection in the south of Anhui province for a long time. However, genome variation, genetic relationships with other pig breeds, and domestication, remain poorly understood. Here, we focus on elucidating the genetic characteristics of the Wannan Black pig and identifying selection signatures during domestication and breeding. Methods: We identified the whole-genome variation in the Wannan Black pig and performed population admixture analyses to determine genetic relationships with other domesticated pig breeds and wild boars. Then, we identified the selection signatures between the Wannan Black pig and Asian wild boars in 100-kb windows sliding in 10 kb steps by using two approaches: the fixation index (F_ST) and π ratios. Results: Resequencing the Wannan Black pig genome yielded 501.52 G of raw data. After calling single-nucleotide variants (SNVs) and insertions/deletions (InDels), we identified 21,316,754 SNVs and 5,067,206 InDels (2,898,582 inserts and 2,168,624 deletions). Additionally, we found genes associated with growth, immunity, and digestive functions. Conclusion: Our findings help in explaining the unique genetic and phenotypic characteristics of Wannan Black pigs, which in turn can be informative for future breeding programs of Wannan Black pigs.

• Journal of Plant Biotechnology
• /
• v.31 no.4
• /
• pp.267-271
• /
• 2004

Transformed sweetpotato (Ipomoea batatas (L.) Lam. cv. Yulmi) plants were developed from embryogenic calli following Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105/pCAMBIA2301 harboring genes for intron $\beta$-glucuronidase (GUS) and kanamycin resistance. Transient expression of GUS gene was found to be higher when embryogenic calli were co-cultivated with Agrobacterium for 2 days. The co-cultured embryogenic calli transferred to selective MS medium containing 1mg/L 2,4-D, 100mg/L kanamycin, and 400mg/L claforan. These embryogenic calli were subcultured to the same selection medium at 4 weeks interval. Kanamycin-resistant calli transferred to hormone-free MS medium with kanamycin gave rise to somatic embryos and then converted into plantlets in the same medium. Southern blot analysis confirmed that the GUS gene was inserted into the genome of the sweetpotato plants. A histochemical assay revealed that the GUS gene was preferentially expressed in the leaf, petiole, and vascular tissue and tip of root.

• Pediatric Infection and Vaccine
• /
• v.23 no.1
• /
• pp.62-66
• /
• 2016

Skin and soft tissue infections (SSTIs) caused by community-associated (CA)-methicillin-resistant Staphylococcus aureus (MRSA) have become a worldwide concern. An otherwise healthy 16-month-old Korean girl was admitted because of skin abscess on the left chest wall with a history of recurrent SSTIs since the age of 6 months. Immunologic evaluation including serum immunoglobulin level and nitroblue-tetrazolium (NBT) test were normal. Pus and nasal swab cultures revealed CA-MRSA ST714-SCCmec type IV with the Panton-Valentine leukocidin (PVL) genes, which was initially reported in the Netherlands in 2006 and has not been previously reported in Korea. The skin abscesses were successfully treated by needle aspiration and the use of antibiotics. In addition, nasal mupirocin was applied as a decolonization method. No more episodes of SSTI were observed over a follow-up period of 10 months.

• Journal of Environmental Health Sciences
• /
• v.39 no.5
• /
• pp.465-473
• /
• 2013

Objectives: This study was undertaken to analyze Staphylococcus aureus from cultivation environments for agricultural products and to confirm antibiotic resistance and enterotoxin genes for the isolated S. aureus. Methods: A total of 648 samples were collected from apple, peach, ginseng and balloon flower farms. S. aureus was isolated from soil, agricultural water, personal hygiene elements (hands, gloves and clothes) and work utensils (boxes). Results: S. aureus was detected in a total of 25 samples and 72 strains were isolated. The resistance rate of the isolated S. aureus strains was confirmed at 33.3%, with 24 resistant strains among the total of 72. Fourteen different patterns types were found, and three pattern types (NV, OX, VA) were confirmed most frequently. As result of the detection of enterotoxin gene type, four gene types (sea: 1, sed: 4, seg: all isolated S. aureus, sei: all isolated S. aureus) were analyzed among a total of nine types. Conclusions: This study demonstrates that personal hygiene techniques should be properly managed, such as washing and sterilization before or after work, because agricultural contamination by S. aureus frequently developed through improper management.

• Journal of Applied Biological Chemistry
• /
• v.60 no.2
• /
• pp.113-117
• /
• 2017

Bioremediation is a technique using microorganisms to clean up contaminated pollutants including heavy metals. It is well known that yeasts have a high capacity to remove a wide range of metals by biosorption. Therefore, this study was focused on to obtain yeast mutant that has strong tolerance to zinc (Zn), one of representative heavy metals. The Zn resistant yeast mutant (ZnR) was induced and isolated by growing yeast cells in media containing 1 mM $ZnCl_2$ and gradually increasing the concentration until 80 mM $ZnCl_2$, in which cells were adapted and survived. The induced ZnR cells showed strong tolerance to Zn stress compared with control cells. Moreover, the ZnR cells showed increased tolerance to cadmium and nickel stress but decreased tolerance to copper stress. The increased tolerance of ZnR cells to Zn stress was due to mutation of genes. This study can be useful in bioremediation of heavy metals as the metal tolerant microorganism was artificially induced in short time.

• Journal of Microbiology and Biotechnology
• /
• v.23 no.8
• /
• pp.1099-1106
• /
• 2013

Tuta absoluta (Povolny, 1994) is a devastating moth to the Solanaceae plants. It is a challenging pest to control, especially on tomatoes. In this work, we studied the entomopathogenic activity of the Cry-forming ${\delta}$-endotoxins produced by Bacillus thuringiensis strain KS and B. thuringiensis kurstaki reference strain HD1 against T. absoluta. These strains carried the cry2, cry1Ab, cry1Aa/cry1Ac, and cry1I genes, and KS also carried a cry1C gene. The ${\delta}$-endotoxins of KS were approximately twofold more toxic against the third instar larvae than those of HD1, as they showed lower 50% and 90% lethal concentrations (0.80 and 2.70 ${\mu}g/cm^2$ (${\delta}$-endotoxins/tomato leaf)) compared with those of HD1 (1.70 and 4.50 ${\mu}g/cm^2$) (p < 0.05). Additionally, the larvae protease extract showed at least six caseinolytic activities, which activated the KS and HD1 ${\delta}$-endotoxins, yielding the active toxins of about 65 kDa and the protease-resistant core of about 58 kDa. Moreover, the histopathological effects of KS and HD1 ${\delta}$-endotoxins on the larvae midgut consisted of an apical columnar cell vacuolization, microvillus damage, and epithelial cell disruption. These results showed that the KS strain could be a candidate for T. absoluta control.

• Microbiology and Biotechnology Letters
• /
• v.19 no.1
• /
• pp.82-87
• /
• 1991

Xanthomonns curnpestris M12, Xunthornonas sp. M28, Acinetuhucter Iwofz GI, and Klebsiella pneumoniae L25, Pseudomonas rnaltophiliu N246 were screened to increase the ability for crude oil utilization. All of these could utilize hexadecane and octane with the exception of N246 strain for only octane biodegradation. Thus N246, M12, and M28, strains were specially examined for octane oxidation. Octane biodegradation by three strains showed the optimal conditions at $30^{\circ}C$, pH 7.0~9.0, and 0.2~0.3% octane concentration as a substrate. It was found that P. multofihila N246 and X. curnpestns M12 had plasmid and the cured plasmid from N246 strain lost octane uitilization. Therefore, it was confirmed that certain genes for octane utilization were Iocated on OCT plasmid in N246 strain. The size of OCT plasmid in N246 strain was 118 kb. The N246 strain was resistant to ampicillin.