20933542
not annotated - annotated - LINNAEUS only
The rapid production of high-titer porcine endogenous retrovirus(PERV)-B env pseudotype and construction of an EGFP-expressing replication competent PERV-A vector.
Porcine endogenous retroviruses (PERVs) present a unique concern associated with xenotransplantation because they have been shown to infect certain human cells in vitro and it is also difficult to generate herds of pigs free of PERVs. A simple system for the production of high-titer MoMLV-PERV pseudotypes is reported; an EGFP-expressing replication-competent molecular clone that allows direct measurement of titer was also constructed. To improve the MLV-based retroviral vector system, a 2.1-kb PERV-B env product was amplified from PK-15 genomic DNA and cloned into the pCL-Eco retroviral vector. The titer of lacZ (PERV-B) from the 293 cells was about 1.0x10(4) CFU/ml. In contrast, the titer of lacZ (PERV-B) from a conventional murine retroviral vector (split genome) was found to be 1.2x10(2) CFU/ml when the PERV-B env expression vector was transfected into TELCeB6 cells, which harbor MFGnlslacZ and the gag-pol-expressing vector. In addition, an infectious PERV-A clone containing enhanced GFP (EGFP) by using a PCR-based method was developed. This EGFP-expressing PERV-A-IRES-EGFP molecular clone was found to be stable genetically on transfection in 293 cells.
Ann file
T1 Species 35 72 porcine endogenous retrovirus(PERV)-B
T2 Species 166 197 Porcine endogenous retroviruses
N2 Reference T2 Taxonomy:369960
T3 Species 199 204 PERVs
N3 Reference T3 Taxonomy:369960
T4 Species 314 319 human
N4 Reference T4 Taxonomy:9606
T5 Species 381 385 pigs
N5 Reference T5 Taxonomy:9823
T6 Species 394 399 PERVs
N6 Reference T6 Taxonomy:369960
T7 Species 630 640 retroviral
N7 Reference T7 Taxonomy:31931
T8 Species 665 671 PERV-B
N8 Reference T8 Taxonomy:369960
T9 Species 787 793 PERV-B
N9 Reference T9 Taxonomy:369960
T10 Species 909 919 retroviral
N10 Reference T10 Taxonomy:31931
T11 Species 984 990 PERV-B
N11 Reference T11 Taxonomy:369960
T12 Species 1135 1141 PERV-A
N12 Reference T12 Taxonomy:369960
N1 Reference T1 Taxonomy:61673 Porcine endogenous retrovirus
T14 Species 874 880 PERV-B
N14 Reference T14 Taxonomy:369960