2006•PubMedRequires access

[Identification of glial cell line-derived neurotrophic factor recombinant retroviral vector and establishment of its packaging cell line PA317].

Wenjie Zhang, Bingcang Li, Xianjun Ren

Abstract

OBJECTIVE: To identify glial cell line-derived neurotrophic factor (GDNF) recombinant retroviral vector and to establish its packaging cell line PA317. METHODS: PA317 cells were transfected with recombinant retroviral vector pLXSN-GDNF using liposomes. The recombinant retroviral particles were then harvested from culture media of G418 resistant transfected cells and analyzed using RT-PCR. Virus titers in supernatants were investigated. RESULTS: Sequencing date indicated that GDNF gene was exactly identical to the sequence in the GeneBank. PA317 cells were transfected with recombinant retroviral vector pLXSN-GDNF using liposomes, and virus titers in supernatants harvested from culture media of (G418 resistant transfected cells were 10(4)10(5) CFU/ml. CONCLUSION: Packaging cell line PA317/pLXSN-GDNF was established.

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Key concepts: Glial cell line-derived neurotrophic factor, Transfection, Viral vector, Cell culture, Recombinant DNA, Biology, Virology, Molecular biology