Rapid assessment of protein-tyrosine phosphatase expression levels by RT-PCR with degenerate primers.

Abstract:

:Using degenerate oligodeoxynucleotide primers we previously obtained cDNA fragments from ten different murine protein-tyrosine phosphatases (PTPases). Employing this same primer set, a method was developed to assess the expression levels of these PTPase family members in a fast and simple way. RT-PCR products of several cell types and tissue samples were used as probes on dot-blots containing the ten different PTPase fragments in equimolar amounts. Hybridization intensities at the various dots reflect the relative expression levels of the corresponding PTPases in the starting material. In this way expression of PTPases during mouse brain development could be monitored. Expression of PTP delta was found to be absent in embryonic stem cells but high in fetal and adult brain. PTP epsilon expression is shown to gradually increase in brain during maturation. Our method is generally applicable to gene families of which the transcripts can be detected with a single degenerate primer pair and is especially useful in situations where only limited amounts of RNA can be obtained.

journal_name

Mol Biol Rep

authors

Hendriks W,Brugman C,Schepens J,Wieringa B

doi

10.1007/BF00997155

subject

Has Abstract

pub_date

1994-03-01 00:00:00

pages

105-8

issue

2

eissn

0301-4851

issn

1573-4978

journal_volume

19

pub_type

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