Application of real-time quantitative PCR in selection of transfected cell strains for transgenic overexpression / 中国实验血液学杂志
Journal of Experimental Hematology
; (6): 1062-1066, 2005.
Article
in Zh
| WPRIM
| ID: wpr-343826
Responsible library:
WPRO
ABSTRACT
To explore the feasibility of real-time quantitative PCR (QRT-PCR) for selecting cell strains which overexpress a certain transgene, expression level of RbAp46 was detected in transfected cell strains by using optimal real-time PCR with SYBR Green I. Meanwhile, semi-quantitative RT-PCR and Western blot were performed to compare with the QRT-PCR. The results showed that values of RbAp46(N) were 2064.42 +/- 253.47, 860.94 +/- 291.07, 234.456 +/- 31.08, 18.17 +/- 5.14 and 1.46 +/- 0.54 in K562/RbAp46, K562/CMV, SHG44/RbAp46 monoclone, SHG44/RbAp46 multiclone and SHG44/CMV, respectively. The results were consistent with that determined by semi-quantitative RT-PCR and Western blot. It is concluded that QRT-PCR provides a highly efficient and reproducible method for selection of transfected cell subclones at different level of transgene expression.
Full text:
1
Database:
WPRIM
Main subject:
Organic Chemicals
/
RNA, Neoplasm
/
Nuclear Proteins
/
Transfection
/
Carrier Proteins
/
Gene Expression Regulation, Neoplastic
/
Chemistry
/
Blotting, Western
/
Transgenes
/
K562 Cells
Limits:
Humans
Language:
Zh
Journal:
Journal of Experimental Hematology
Year:
2005
Document type:
Article