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The resultant replication-competent molecular clone was designated as p05MYKL045.1.
Each molecular clone was transfected into COS-7 cells.
The NL4-3-based molecular clone was constructed by replacing the pol-coding region with the HIV-1 BH10 strain.
To confirm this, a HIVΔvif molecular clone was transfected into QT6 cells and the resulting progeny used to infect the CEMX174 cell line.
To generate a more replication-fit virus, an infectious molecular clone was generated by directly cloning a virus from a SHIV-1157i-infected RM after it had developed AIDS 2.7 years post-inoculation.
The mt molecular clone was obtained by insertion of the XhoI-HindIII fragment of pBlue3'LTR into the 3'LTR of the HIV-1 molecular clone pLAI and the ClaI-XbaI fragment of pBlue5'LTR into the 5'LTR of the HIV-1 molecular clone pLAI.
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A selected molecular clone is tested in a single round assay with luciferase readout that results in short-term assays with high reproducibility and sensitivity.
Because direct sequencing of the ITS PCR products of the new species resulted in DNA sequences with overlapping signals, molecular cloning was performed.
Molecular cloning was performed according to standard protocols.
Molecular cloning was conducted using E. coli NEB-10-Beta (New England Biolabs, NEB).
Molecular cloning was done on the subtracted DNAs and the positive isolates were extracted, purified and sequenced for analysis.
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