[Detection of p27/kip1 mRNA in blood cells by nonradioactive ribonuclease protection assay]

Rinsho Byori. 1996 May;44(5):483-6.
[Article in Japanese]

Abstract

Cyclin and cyclin-dependent kinase(cdk) complexes, and their inhibitors (CKIs) play important roles in growth regulation on the cells. p27/kip1 is a CKI associated with G1 arrest induced by cell to cell contact, transforming growth factor-beta and cyclic AMP. The abnormality of p27/Kip1 genes in human tumors usually appears as a steady level defect of expression, since mutations in them is rare. Thus it is important to estimate the expression level of this gene. To detect the change of p27/Kip1 mRNA level in blood cells, we developed the ribonuclease protection assay using nonradioactive riboprobe which was produced by reverse transcriptase-polymerase chain reaction (RT-PCR) with T7 promoter-added antisense primer and the in vitro transcription system. Our assay may be useful for clinical evaluation of the mRNA level.

Publication types

  • English Abstract

MeSH terms

  • Base Sequence
  • Cell Cycle Proteins*
  • Cyclin-Dependent Kinase Inhibitor p27
  • Humans
  • Microtubule-Associated Proteins / genetics*
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Polymorphism, Single-Stranded Conformational
  • RNA, Messenger / blood*
  • Ribonucleases
  • Tumor Suppressor Proteins*

Substances

  • Cell Cycle Proteins
  • Microtubule-Associated Proteins
  • RNA, Messenger
  • Tumor Suppressor Proteins
  • Cyclin-Dependent Kinase Inhibitor p27
  • Ribonucleases