Improved PCR-Based Subtractive Hybridization Strategy for Cloning Differentially Expressed Genes
Citations Over Time
Abstract
An improved PCR-based subtractive hybridization strategy was used to clone apoptosis-related genes induced by all-trans retinoic acid (ATRA) from human promyelocytic leukemia cell line HL-60 cells. The protocol used the cap-finder method, long-distance PCR, streptavidin magnetic bead-mediated subtraction and spin column chromatography. Twenty-seven clones related to apoptosis were identified by reverse dot blot assay. Seventeen were known genes, of which seven have been reported to be apoptosis related. The remaining 10 were unknown genes, five of which were sequenced and named apr-1 to apr-5. apr-1, apr-2, apr-3 and TNF were reidentified by reverse dot blot, and it is suggested that they might be related to apoptosis. The results suggest that this strategy might be efficient for large-scale cloning of differentially expressed genes in target cells.
Related Papers
- → Comparison of dot-blot and northern blot hybridizations in the determination of genetic relatedness of United States bluetongue virus serotypes(1988)7 cited
- → QUANTITATION OF mRNA SPECIFIC FOR HEAVY AND LIGHT CHAINS OF IgG IN HYBRIDOMAS DURING DIFFERENT PHASES OF BATCH CULTURE(1991)6 cited
- Construction of differential expression library for survival shrimps infected with TSV and uninfected shrimps.(2010)
- Screening differentially expressed genes from suppression subtractive hybridizational positive clones using reverse mRNA dot blot technique(2002)
- A rapid method for subtractive screening of hepatoma apoptotic cells cDNA library(1999)