pLXSN
6 kB
22 Expression in Eukaryotic Cells: Gene Transfer Using Retroviral Vectors
339
[I] Long Terminal Repeat (LTR)
0
Packaging Signal
II- SV 40 Promoter Enhancer, Early Region
Fig. 1. Retroviral Vector pLXSN
• Competent bacteria used to replicate the plasmid
• A packaging cellline for the production of replication-incompetent ecotropic retroviral particles. These can be used to infect rodent cells. For the
production of amphotropic replication-incompetent viruses that have a
broad host specificity a second packaging cellline must be infected with
the ecotropic viruses.
• The target cell
Subprotocol 1
Synthesis of the Insert
Procedure
The DNA that is tobe inserted into the genome ofthe target cells is usually
synthesized by PCR (see Chapter 14). Suitable restriction sites to facilitate
the ligation of the DNA into the vector need tobe considered when designing the appropriate PCR primers. If possible, we preferentially include an
6 kB
22 Expression in Eukaryotic Cells: Gene Transfer Using Retroviral Vectors
339
[I] Long Terminal Repeat (LTR)
0
Packaging Signal
II- SV 40 Promoter Enhancer, Early Region
Fig. 1. Retroviral Vector pLXSN
• Competent bacteria used to replicate the plasmid
• A packaging cellline for the production of replication-incompetent ecotropic retroviral particles. These can be used to infect rodent cells. For the
production of amphotropic replication-incompetent viruses that have a
broad host specificity a second packaging cellline must be infected with
the ecotropic viruses.
• The target cell
Subprotocol 1
Synthesis of the Insert
Procedure
The DNA that is tobe inserted into the genome ofthe target cells is usually
synthesized by PCR (see Chapter 14). Suitable restriction sites to facilitate
the ligation of the DNA into the vector need tobe considered when designing the appropriate PCR primers. If possible, we preferentially include an