• Title/Summary/Keyword: electroportion

Search Result 2, Processing Time 0.009 seconds

Transformation of Bacillus brevis P176-2 with Plasmid DNA by Electroporation (전기천공법에 의한 Bacillus brevis P176-2의 형질전환)

  • 채기수;엄경일
    • The Korean Journal of Food And Nutrition
    • /
    • v.5 no.2
    • /
    • pp.77-83
    • /
    • 1992
  • The optimum conditions and mechanisms for the plasmid-mediated genetic transformation of intact cells of Bacillus brevis Pl76-2, an extracellular protein producing bacterium by electroporation were investigated. It was found that pUB110 Plasmid DNA can be introduced into intact bacterial cells by electroporation. The frequency of transformation by this electroporation system depended upon the initial electric field strength, the capacity of the electric discharge capacitor, growth stage, number of successive pulses and composition of electroporation buffer. It was effective for transformation that cells were harvested, washed and resuspended with HSM [7M HEPES(PH 7.4), 272mM sucrose, 1 mM MgCl2] electroporation buffer when cell growth was attained to 1.2 at OD660. A maximum frequency of transformation of 2.40$\times$104 transformants per$\mu$g plasmid DNA was obtained by two succesive Pulses with an initial electric field strength of 12.5kV/cm and with a capacitance of 7.3uF.

  • PDF

Prolonged Gene Expression Following Erythrocyte-Mediated Delivery of TRAIL Plasmid DNA (혈구세포 수송체로 투여된 트레일 유전자의 혈중 발현 지속 효과)

  • Byun, Hyang-Min;Kwon, Kyoung-Ae;Shin, Jee-Young;Oh, Yu-Kyoung
    • Journal of Pharmaceutical Investigation
    • /
    • v.33 no.4
    • /
    • pp.261-265
    • /
    • 2003
  • Tumor necrosis facto-related apoptosis-inducing ligand (TRAIL) is a recently identified member of the tumor necrosis factor cytokine superfamily. TRAIL has been shown to induce apoptosis in a number of tumor cells whereas cells from most of normal tissues are highly resistant to TRAIL-induced apoptosis. These observations have raised considerable interest in the use of TRAIL in tumor therapy. In this study we report the biodistribution fates and serum expression pattern of plasmid DNA encoding TRAIL (pTRAIL) delivered in erythrocyte ghosts (EG). pTRAIL was loaded into EG by electroportion in a hypotonic medium The mRNA expression of pTRAIL was prolonged following delivery in EG-encapsulated forms. EG containing pTRAIL showed significant levels of mRNA expression in the blood over 9 days. The organ expression patterns of pTRAIL delivered via EG, however, did not significantly differ from those of naked pTRAIL, indicating that the expression-enhancing effect of EG containing pTRAIL was localized to the blood. These results suggest that pTRAIL-loaded EG might be of potential use in the treatment of hematological diseases such as TRAIL-sensitive leukemia.