PMID- 12850372
OWN - NLM
STAT- MEDLINE
DCOM- 20030812
LR  - 20190816
IS  - 0165-4608 (Print)
IS  - 0165-4608 (Linking)
VI  - 144
IP  - 2
DP  - 2003 Jul 15
TI  - Localization of transfected B7-1 (CD80) DNA in human melanoma cells after 
      particle-mediated gene transfer.
PG  - 106-11
AB  - The purpose of this study was to evaluate stable DNA transfection of M-21 human 
      melanoma cells with particle-mediated gene transfer (PMGT) with B7-1 cDNA and to 
      identify sites of gene integration. Stable B7-1 transfectants (M-21-B7) were 
      obtained with PMGT using a plasmid vector containing cDNA for both B7-1 and 
      neomycin phosphotransferase, with subsequent selection with G418. The transfected 
      cells were flow sorted by B7-1 expression into two populations, bright and dim. 
      The bright population had 85%-90% of cells expressing B7-1; the dim population 
      had less than 50% of cells with B7-1 expression. Chromosome analysis with 
      fluorescence in situ hybridization (FISH) and G-banding showed that 70% of bright 
      cells had two main integration sites, with extensive amplification of the 
      transgene. The dim population had random signal distribution, with little or no 
      amplification, despite G418 selection. Because B7-1 has been mapped to 3q21, FISH 
      was performed using a chromosome 3 painting probe (WCP) together with a probe for 
      B7-1. In transfected bright M-21 cells, amplified genes that hybridized with the 
      B7-1 construct were localized to chromosome 3 material inserted into marker 
      chromosomes. These data suggest that B7-1 insertion may involve homologous 
      recombination, but maintenance of integration and amplification required 
      selection.
FAU - McCarthy, Donna O
AU  - McCarthy DO
AD  - Comprehensive Cancer Center, University of Wisconsin Hospital and Clinics, 
      Madison, WI 53792, USA.
FAU - Meisner, Lorraine F
AU  - Meisner LF
FAU - Bourdeau-Heller, Jeanne
AU  - Bourdeau-Heller J
FAU - Roberts, Timothy R
AU  - Roberts TR
FAU - Wu, Shi Qi
AU  - Wu SQ
FAU - Warner, Thomas F
AU  - Warner TF
FAU - Albertini, Mark R
AU  - Albertini MR
LA  - eng
GR  - CA68466/CA/NCI NIH HHS/United States
GR  - NR00088/NR/NINR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Cancer Genet Cytogenet
JT  - Cancer genetics and cytogenetics
JID - 7909240
RN  - 0 (B7-1 Antigen)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - B7-1 Antigen/*genetics
MH  - DNA/*analysis
MH  - *Gene Transfer, Horizontal
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Melanoma/*genetics
MH  - Plasmids
MH  - Transfection
MH  - Transgenes
MH  - Tumor Cells, Cultured
EDAT- 2003/07/10 05:00
MHDA- 2003/08/13 05:00
CRDT- 2003/07/10 05:00
PHST- 2003/07/10 05:00 [pubmed]
PHST- 2003/08/13 05:00 [medline]
PHST- 2003/07/10 05:00 [entrez]
AID - S0165460802009408 [pii]
AID - 10.1016/s0165-4608(02)00940-8 [doi]
PST - ppublish
SO  - Cancer Genet Cytogenet. 2003 Jul 15;144(2):106-11. doi: 
      10.1016/s0165-4608(02)00940-8.