The following plasmids were generated by mutagenesis: pJS82 [p1D4-hrGFP II-PCMV: :hrAir-ccapr1-Like[DELTA]105K132G; PCMV
: :gfp], pJS86 [p1D4-hrGFP II-PCMV: :hrAir-aarI-Like[DELTA]58K113A; PCMV
: :gfp], and pJS88 [p1D4-hrGFP II-PCMV: :hrAir-aar1-Like[DELTA]58K205G; PCMV
Culture medium (100 f l) from C6 cultures transfected with pCMV, pCMV-[SOD1.sup.wt], or pCMV-[SOD1.sup.G93A] was collected and measured for S100B content using S100B human ELISA kit (BioVendor) following the manufacturer's instructions.
C6 cells were transiently transfected with pCMV (mock), pCMV-[SOD1.sup.wild-type] ([SOD1.sup.wt]), or pCMV-[SOD1.sup.G93A] ([SOD1.sup.G93A]).
All tissues were negative for the presence of PCMV, PLHV, PERV, sHEV1 and sHEV2, and sEMCV.
Porcine reproductive and VDL RT-PCR respiratory syndrome virus (PRRSV) Swine influenza virus VDL RT-PCR (SIV) Porcine endogenous VDL RT-PCR retrovirus (PERV) Porcine enterovirus (PEV) VDL RT-PCR Porcine respiratory VDL RT-PCR (Qiagen 283615) corona virus (PRCV) Transmissible VDL RT-PCR (Qiagen 283615) gastroenteritis virus (TGEV) Porcine circovirus types VDL Multiplex PCR 1 and 2 (PCV) Porcine lymphotropic VDL PCR herpes virus type-1 (PLHV-1) Swine hemagglutinating VDL RT-PCR encephalomyelitis virus (sHEV) Porcine parvovirus (PPV) VDL PCR Porcine cytomegalovirus VDL PCR (PCMV) Mycoplasma hyopneumoniae VDL PCR Pseudorabies virus (PRV) VDL Virus isolation Encephalomyocarditis VDL Virus isolation virus (EMCV) Rotavirus (type A) VDL Antigen capture ELISA Chlamydia spp.
For luciferase vector transfection, pCMV
[beta]-galactosidase plasmid (Clontech, USA) was used for normalization.
Glass, University of California San Diego) or a control vector (pCMV
) was cotransfected with the luciferase reporter plasmid.
A total of 25 x [10.sup.6] cells were cotransfected for 20 min in 0.5 mL STBS containing 200 [micro]g DEAE- dextran and 0.5 [micro]g plasmid to be tested together with 0.5 [micro]g of the pCMV
plasmid expressing [beta]-galactosidase under the control of the CMV promoter (pCMV-[beta]-gal).