PMID- 8391178
OWN - NLM
STAT- MEDLINE
DCOM- 19930723
LR  - 20081121
IS  - 0042-6822 (Print)
IS  - 0042-6822 (Linking)
VI  - 195
IP  - 1
DP  - 1993 Jul
TI  - Factors affecting retroviral vector function and structural integrity.
PG  - 1-5
AB  - Recombinant retroviruses are widely used for gene transfer into eukaryotic cells 
      and exhibit significant potential for human gene therapy. Despite the utility of 
      retroviral vectors, their design is still essentially empirical. We have 
      constructed a series of reciprocal, double-gene vectors to compare the dual 
      expression of beta-galactosidase (beta-gal) and neomycin phosphotransferase 
      (neor) in a retroviral delivery system. The first gene of the pair was driven by 
      the viral LTR promoter and the internal gene was regulated by either the SV40 
      virus early promoter or the cytomegalovirus (CMV) major late promoter. Clones of 
      vector producer cells were isolated either by G418 selection for expression of 
      neor, or by fluorescence-activated cell sorting for expression of beta-gal, and 
      the activity of both genes was evaluated. In general, vectors using the SV40 
      promoter performed better than those with the CMV promoter, regardless of whether 
      the selected gene was regulated by the LTR or the internal promoter. Southern 
      analysis of clones indicated that loss of beta-gal gene function was related to 
      significant rearrangements and deletions in vector structure. We also found that 
      the arrangement of genes within the vector was important. When beta-gal preceded 
      neor, gene expression and vector stability were markedly enhanced relative to 
      vectors containing these genes in the inverse order.
FAU - McLachlin, J R
AU  - McLachlin JR
AD  - Genetic Therapy, Inc., Gaithersburg, Maryland 20878.
FAU - Mittereder, N
AU  - Mittereder N
FAU - Daucher, M B
AU  - Daucher MB
FAU - Kadan, M
AU  - Kadan M
FAU - Eglitis, M A
AU  - Eglitis MA
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Virology
JT  - Virology
JID - 0110674
SB  - IM
MH  - Blotting, Southern
MH  - Cloning, Molecular/methods
MH  - Cytomegalovirus/genetics
MH  - *Genetic Vectors
MH  - Promoter Regions, Genetic
MH  - Retroviridae/*genetics
MH  - Simian virus 40/genetics
MH  - Transduction, Genetic
EDAT- 1993/07/01 00:00
MHDA- 1993/07/01 00:01
CRDT- 1993/07/01 00:00
PHST- 1993/07/01 00:00 [pubmed]
PHST- 1993/07/01 00:01 [medline]
PHST- 1993/07/01 00:00 [entrez]
AID - S0042-6822(83)71340-1 [pii]
AID - 10.1006/viro.1993.1340 [doi]
PST - ppublish
SO  - Virology. 1993 Jul;195(1):1-5. doi: 10.1006/viro.1993.1340.