Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1997 Jun 1;25(11):2233–2235. doi: 10.1093/nar/25.11.2233

Direct identification of differentially expressed genes by cycle sequencing and cycle labelling using the differential display PCR primers.

M Buess 1, C Moroni 1, H H Hirsch 1
PMCID: PMC146722  PMID: 9153328

Abstract

Differential display PCR (DD-PCR) is an mRNA fingerprinting technique to identify differentially expressed genes by comparative display of arbitrarily amplified cDNA subsets. This attractively simple screening method was, however, followed by a labour intensive multistep identification procedure for DD-PCR products. In this report we demonstrate for the mouse mast cell protease 2 (MMCP-2) and the cytotoxic T-lymphocyte associated gene transcript CTLA-1 a streamlined approach by (i) direct cycle sequencing with the upstream differential display (DD) primer, followed by (ii) the PCR based generation of an antisense northern probe with the downstream anchor primer.

Full Text

The Full Text of this article is available as a PDF (93.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bauer D., Müller H., Reich J., Riedel H., Ahrenkiel V., Warthoe P., Strauss M. Identification of differentially expressed mRNA species by an improved display technique (DDRT-PCR). Nucleic Acids Res. 1993 Sep 11;21(18):4272–4280. doi: 10.1093/nar/21.18.4272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brunet J. F., Denizot F., Suzan M., Haas W., Mencia-Huerta J. M., Berke G., Luciani M. F., Golstein P. CTLA-1 and CTLA-3 serine esterase transcripts are detected mostly in cytotoxic T cells, but not only and not always. J Immunol. 1987 Jun 15;138(12):4102–4105. [PubMed] [Google Scholar]
  3. Gough N. M. Rapid and quantitative preparation of cytoplasmic RNA from small numbers of cells. Anal Biochem. 1988 Aug 15;173(1):93–95. doi: 10.1016/0003-2697(88)90164-9. [DOI] [PubMed] [Google Scholar]
  4. Hirsch H. H., Nair A. P., Backenstoss V., Moroni C. Interleukin-3 mRNA stabilization by a trans-acting mechanism in autocrine tumors lacking interleukin-3 gene rearrangements. J Biol Chem. 1995 Sep 1;270(35):20629–20635. doi: 10.1074/jbc.270.35.20629. [DOI] [PubMed] [Google Scholar]
  5. Hirsch H. H., Nair A. P., Moroni C. Suppressible and nonsuppressible autocrine mast cell tumors are distinguished by insertion of an endogenous retroviral element (IAP) into the interleukin 3 gene. J Exp Med. 1993 Aug 1;178(2):403–411. doi: 10.1084/jem.178.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Liang P., Averboukh L., Pardee A. B. Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization. Nucleic Acids Res. 1993 Jul 11;21(14):3269–3275. doi: 10.1093/nar/21.14.3269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Liang P., Pardee A. B. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science. 1992 Aug 14;257(5072):967–971. doi: 10.1126/science.1354393. [DOI] [PubMed] [Google Scholar]
  8. Nair A. P., Diamantis I. D., Conscience J. F., Kindler V., Hofer P., Moroni C. A v-H-ras-dependent hemopoietic tumor model involving progression from a clonal stage of transformation competence to autocrine interleukin 3 production. Mol Cell Biol. 1989 Mar;9(3):1183–1190. doi: 10.1128/mcb.9.3.1183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Serafin W. E., Reynolds D. S., Rogelj S., Lane W. S., Conder G. A., Johnson S. S., Austen K. F., Stevens R. L. Identification and molecular cloning of a novel mouse mucosal mast cell serine protease. J Biol Chem. 1990 Jan 5;265(1):423–429. [PubMed] [Google Scholar]
  10. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Wang X., Feuerstein G. Z. Direct sequencing of DNA isolated from mRNA differential display. Biotechniques. 1995 Mar;18(3):448–453. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES