• Title/Summary/Keyword: kanamycin-resistant gene

Search Result 75, Processing Time 0.022 seconds

Characterization and Methanol Biosynthesis of a Methane-Oxidizing Bacterium, Methylomonas sp. SM4, Isolated from Rice Paddy Field Soil (논에서 분리한 메탄산화세균 Methylomonas sp. SM4의 특성과 메탄올 생합성)

  • Park, Sung Min;Madhavaraj, Lavanya;Kim, Si Wouk
    • KSBB Journal
    • /
    • v.32 no.2
    • /
    • pp.124-132
    • /
    • 2017
  • A methane-oxidizing bacterium was isolated from rice paddy field soil around Jeollanam-do province, Korea, and characterized. The isolate was gram-negative, orange pigmented and short rod ($1.1-1.2{\times}1.6-1.9{\mu}m$). It was catalase and urease-negative but oxidase-positive. The strain utilized methane and methanol as sole carbon and energy sources. It had an ability to grow with an optimum pH 7.0 and an optimum growth temperature $30^{\circ}C$. The strain was resistant to antibiotic polymyxin B but sensitive to streptomycin, kanamycin, ampicillin, chloramphenicol and rifampicin. The isolate required copper for their growth with concentration range of $2-25{\mu}M$, with an optimum of $10{\mu}M$. Under optimal culture condition, specific cell growth rate and generation time were found to be $0.046hr^{-1}$ and 15.13 hr, respectively. Phylogenetic analysis based on 16S rDNA sequences indicated that the strain formed a tight phylogenetic lineage with Methylomonas koyamae with a value of 99.4% gene sequence homology. So, we named the isolate as Methylomonas sp. SM4. 8.6 mM methanol was accumulated in the reaction mixture containing 70 mM sodium formate and 40 mM $MgCl_2$ (MDH inhibitor) under atmosphere of methane:air (40:60) mixture for 24 hr at $30^{\circ}C$.

Probiotic Properties of Pediococcus pentosaceus SH-10 Isolated from the Hard Clam Meretrix meretrix Shikhae (백합(Meretrix meretrix)식해에서 분리한 Pediococcus pentosaceus SH-10의 생균제적 특성)

  • Song, Hyun-Jung;Kim, Kang-Jin;Kim, Hee-Dai;Yoo, Jung-Hee;Koo, Jae-Geun;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.44 no.6
    • /
    • pp.605-611
    • /
    • 2011
  • This study examined the suitability of characteristics of potential strains of probiotic bacteria. Among 25 lactic acid bacteria isolated from Korean traditional fermented food, the Hard Clam Meretrix meretrix Shikhae, the SH-10 strain, which exhibited superior resistance to low pH and bile salts, was selected as a potential probiotic bacteria. By examining carbohydrate utilization, morphological properties, and the 16S rRNA gene sequence, the SH-10 strain was identified as Pediococcus pentosaceus (hereafter, P. pentosaceus SH-10). P. pentosaceus SH-10 was resistant to amikacin, cefotetan, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, and vancomycin. Tests of antimicrobial activities against pathogens such as Bacillus cereus, Listeria monocytogenes, Salmonella choleraesuis, and Staphylococcus aureus, indicated that P. pentosaceus SH-10 inhibited the growth of pathogenic bacteria. These results suggest that P. pentosaceus SH-10 can be developed as a probiotic bacteria.

Transformation of Bottle Gourd Rootstock (Lagenaria siceraria Standl.) using GFP gene (GFP유전자를 이용한 대목용 박 형질전환)

  • Lim, Mi-Young;Park, Sang-Mi;Kwon, Jung-Hee;Han, Sang-Lyul;Shin, Yoon-Sup;Han, Jeung-Sul;Harn, Chee-Hark
    • Journal of Plant Biotechnology
    • /
    • v.33 no.1
    • /
    • pp.33-37
    • /
    • 2006
  • Bottle gourd (Lagenaria siceraria Standl.) has been used as a rootstock for the watermelon cultivation because of better growth ability at low temperature and avoidance from contamination of the soil disease. Since the genetic source for the elite rootstock is limited in nature, the genetic engineering method is inevitable to develop new lines especially to obtain the functionally important or multi-disease resistant bottle gourd. Recently, our lab has set up a successful system to transform the bottle gourd. in order to monitor the transformation process, GFP gene is used. Cotyledons of the inbred line 9005, 9006 and G5 were used to induce the shoot under the selection media with MS + 30 g/L sucrose + 3.0 mg/L BAP + 100 mg/L kanamycin + 500 mg/L cefotaxime + 0.5 mg/L $AgNO_3$, pH 5.8. The shoot was developed from the cut side of the explants after 3 weeks on the selection media. The shoot was incubated in the rooting media with 1/2 MS + 30 g/L sucrose + 0.1 mg/L IAA + 50 mg/L kanamycin + 500 mg/L cefotaxime, pH 5.8 and moved to pot for acclimation. Although the shoot development rate was depended on the genotype, the G5 was the best line to be transformed. Monitoring GFP expression from the young shoot under microscope could make the selection much easier to distinguish the transformed shoot from the non-transformed shoots.

Transformation of Gourd through Leaf Explant Regeneration (잎 절편의 재분화에 의한 참박 형질전환)

  • Cho, Song-Mi;Moon, Sun-Jin;Chung, Soo-Jin;Kim, Mi-Seong;Kim, Young-Cheol;Yang, Kwang-Yeol;Choi, Yong-Soo;Sapkota, Kumar;Cho, Baik-Ho;Kim, Kwang-Sang
    • Korean Journal of Plant Resources
    • /
    • v.19 no.5
    • /
    • pp.634-639
    • /
    • 2006
  • In order to develop a disease-resistant root stock for the growth of watermelon, an efficient regeneration system of the gourd(Lagenaria leucantha Duch.) inbred line GO701-2 via organogenesis was established in this experiment. Using proximal parts of cotyledon explant excised from germinated seedling in vitro, maximum adventitious shoot formation (39%) was achieved on MS medium where cytokinin (BA) and auxin (IAA) were added at a concentration of 3mg/L and 0.1mg/L, respectively. Roots of the elongated shoots were successfully formed on MS medium without adding any plant growth regulators. The cucumber CsGolS1 gene known as a resistance gene against biotic and abiotic stresses, was constructed into the binary vector pBI121 under the control of CaMV 35S promoter. When the gene was introduced into the genome of gourd by Agrobacterium-mediated transformation, putative transgenic plants were obtained with the transformation efficiency of approximately 20 percent.

Transformation is Mechanism of Gene Transfer in Soil (토양에서 Transformation에 의한 유전자 전이)

  • ;Stotzky, G.
    • Korean Journal of Microbiology
    • /
    • v.28 no.3
    • /
    • pp.210-218
    • /
    • 1990
  • The survival and transfer of chromosomal genes coding for the synthesis of amino acids (threonine, tryptophan, histidine, leucine, methionine) and of plasmid-borne genes coding for resistance to antibiotics (chloramphenicol, kanamycin, erythromycin) by transformation in sterile and nonsterile soil (the soil was amended to 12% vol/vol with the clay mineral, montmorillonite) was studied. In pure culture, the numbers of vegetative cells of the Bacillus subtilis strains decreased by 1 to 1.5 orders of magnitude within one week, but spores of each strain showed lesser decreases. In sterile soil, the populations of vegetative cells and spores decreased by 1.5 to 3 orders of magnitude within 2 to 4 days and then showed little additional decreased. The transformation frequencies (number of transformants/numbers of donors and recipients) of individual amino acid-genes invitro ranged from $1.3{\pm}0.6{\times}10^{-6}$ to $6.0{\pm}2.36{\times}10^{-6}$, of two amino acid-genes from $8.5{\pm}0.7{\times}10^{-8}$ to $3.1{\pm}0.6{\times}10^{-7}$, and of the antibiotic-resistance genes from $1.5{\pm} 0.2{\itmes} 10^{-7}$ to $1.4{\pm} 0.4{\times} 10^{-5}$ . In sterile soil, the frequencies of transfer of individual amino acid-genes ranged from $2.0{\times} 10^{-7}$ to $2.0{\times} 10^{-5}$ and of the antibiotic-resistance genes from $2.0{\times} 10^{-7}$ to $9.4{\pm} 4.7{\times} 10^{-6}$. The transfer of two amino acid-genes in sterile soil was detected at a frequency of $2.0{\times} 10^{-6}$ to $4.5{\times} 10^{-6}$, but only in three instances. The transformation frequencies of antibiotic-resistance genes in nonsterile soil were essentially similar to those in sterile soil. However, to detect transformants in nonsterile soil, higher concentrations of antibiotics were needed, as the result of the large numbers of indigenous soil bacteria resistant to the concentration of antibiotics used in the sterile soil and in vitro studies. The results of these studies show that genes can be transferred by transformation in soil and that this mechanism of transfer must be considered in risk assessment of the release of genetically engineered microorganisms to the environment.

  • PDF