Figure 5.
Figure 5. T lymphocytes immortalized by HTLVRex- contain the expected Rex- mutation and express tax/rex mRNA. (A) HTLV-1 genome fragment containing the Rex start codon was amplified by PCR from high molecular weight DNA of wtHTLV-1 and HTLVRex- immortalized cells. PCR-amplified product was incubated in the presence or absence of SphI, separated on 2% agarose gel, and visualized by ethidium bromide staining. (B) Detection of tax/rex mRNA in the wtHTLV-1 and HTLVRex- immortalized PBMCs. Total RNA was prepared from fresh PBMCs, wtHTLV-1, and HTLVRex- immortalized PBMCs. Approximately 0.3 μg RNA was subjected to a coupled 40-cycle RT-PCR in the presence (+) or absence (-) of reverse transcriptase (RT). Following RT-PCR, 25-cycle nested PCR was performed. PCR product was separated on 2% agarose gel and visualized by ethidium bromide staining.

T lymphocytes immortalized by HTLVRex- contain the expected Rex- mutation and express tax/rex mRNA. (A) HTLV-1 genome fragment containing the Rex start codon was amplified by PCR from high molecular weight DNA of wtHTLV-1 and HTLVRex- immortalized cells. PCR-amplified product was incubated in the presence or absence of SphI, separated on 2% agarose gel, and visualized by ethidium bromide staining. (B) Detection of tax/rex mRNA in the wtHTLV-1 and HTLVRex- immortalized PBMCs. Total RNA was prepared from fresh PBMCs, wtHTLV-1, and HTLVRex- immortalized PBMCs. Approximately 0.3 μg RNA was subjected to a coupled 40-cycle RT-PCR in the presence (+) or absence (-) of reverse transcriptase (RT). Following RT-PCR, 25-cycle nested PCR was performed. PCR product was separated on 2% agarose gel and visualized by ethidium bromide staining.

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