돼지 내인성 레트로바이러스의 복제 및 감염억제를 위한 조절유전자와 RNA 간섭서열의 발현벡터 구축 Construction of vectors containing regulatory genes and RNA interference sequence to inhibit the replication and infectivity of porcine endogenous retrovirus원문보기
Porcine Endogenous Retrovirus (PERV) represent a potential infectious risk in xenotransplantation. Known infectious PERV have been assigned to the PERV γ-1 family, consisting of subfamilies A, B, and C. To measure the inhibitory effect of the replication and infectivity of PERV, in porcine cells (MP...
Porcine Endogenous Retrovirus (PERV) represent a potential infectious risk in xenotransplantation. Known infectious PERV have been assigned to the PERV γ-1 family, consisting of subfamilies A, B, and C. To measure the inhibitory effect of the replication and infectivity of PERV, in porcine cells (MPK) producing A, B, and C subtypes, transformed with the combinated vectors. The new regulatory genes were human APOBEC 3G, Africa green monkey Trim5α,shRNAi/PERVpol69 , and shRNAi/PERVpol760, which of one, two, and four genes were combinated into the viral packaging signal-deleted pLPCX vector under the control of pCMV IE and 5'LTR. The mRNA expression of the new regulatory genes on the trasnfected the MPK cells were detected by RT-PCR. These vectors containing regulatory genes and the RNA interference (RNAi) technique would be useful for the inhibition of the replication and infectivity of PERV. The inhibition of the replication and infectivity of PERV could be confirmedby the quantitative analysis of viral mRNA, proviral genomic DNA, viral RNA and virus particles of MPK cells by PCR and RT-PCR with proper primers of pol and env (A, B, and C) genes.
Porcine Endogenous Retrovirus (PERV) represent a potential infectious risk in xenotransplantation. Known infectious PERV have been assigned to the PERV γ-1 family, consisting of subfamilies A, B, and C. To measure the inhibitory effect of the replication and infectivity of PERV, in porcine cells (MPK) producing A, B, and C subtypes, transformed with the combinated vectors. The new regulatory genes were human APOBEC 3G, Africa green monkey Trim5α,shRNAi/PERVpol69 , and shRNAi/PERVpol760, which of one, two, and four genes were combinated into the viral packaging signal-deleted pLPCX vector under the control of pCMV IE and 5'LTR. The mRNA expression of the new regulatory genes on the trasnfected the MPK cells were detected by RT-PCR. These vectors containing regulatory genes and the RNA interference (RNAi) technique would be useful for the inhibition of the replication and infectivity of PERV. The inhibition of the replication and infectivity of PERV could be confirmedby the quantitative analysis of viral mRNA, proviral genomic DNA, viral RNA and virus particles of MPK cells by PCR and RT-PCR with proper primers of pol and env (A, B, and C) genes.
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