• Title/Summary/Keyword: transducing particles

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Stability of Retroviral Vectors Against Ultracentrifugation Is Determined by the Viral Internal Core and Envelope Proteins Used for Pseudotyping

  • Kim, Soo-hyun;Lim, Kwang-il
    • Molecules and Cells
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    • v.40 no.5
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    • pp.339-345
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    • 2017
  • Retroviral and lentiviral vectors are mostly pseudotyped and often purified and concentrated via ultracentrifugation. In this study, we quantified and compared the stabilities of retroviral [murine leukemia virus (MLV)-based] and lentiviral [human immunodeficiency virus (HIV)-1-based] vectors pseudotyped with relatively mechanically stable envelope proteins, vesicular stomatitis virus glycoproteins (VSVGs), and the influenza virus WSN strain envelope proteins against ultracentrifugation. Lentiviral genomic and functional particles were more stable than the corresponding retroviral particles against ultracentrifugation when pseudotyped with VSVGs. However, both retroviral and lentiviral particles were unstable when pseudotyped with the influenza virus WSN strain envelope proteins. Therefore, the stabilities of pseudotyped retroviral and lentiviral vectors against ultracentrifugation process are a function of not only the type of envelope proteins, but also the type of viral internal core (MLV or HIV-1 core). In addition, the fraction of functional viral particles among genomic viral particles greatly varied at times during packaging, depending on the type of envelope proteins used for pseudotyping and the viral internal core.

Asymmetry of Cotransduction Frequency Occured When the Transposon Is Used as a Marker (전위인자가 표식인자로 사용될때 발생하는 동시형질도입의 비대칭성)

  • 김창훈;박찬규
    • Korean Journal of Microbiology
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    • v.30 no.2
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    • pp.83-87
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    • 1992
  • When the transposon is used as a selectable marker. the cotransduction frequency depends on the selection order of the markers. In this study. we adopted a mathematical approach to explain this phenomenon. At first, the formation of transducing particles were considered in five different configurations. Then the probability functions indicating the possibilities for a marker to be fixed were mathematically formulated on the basis of probability density concept. After actual values and useful assumptions were integrated into the Formula. resulting frequencies from theoretical calculations were compared with actual data. Such analysis let us conclude that the asymmetry in the frequency arose from the lack of homology required for homologous recombination due to the transposon insertion and from the suppression of recombination around the region where the transposon is inserted.

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