[Update on a diagnostic test for choroideremia: the protein truncation test (PTT)].
OBJECTIVE
The aim of this study was to define the RT-PCR-PTT parameters for CHM gene analysis and to evaluate its interest as a method for CHM mutation screening.


METHODS
The entire CHM coding region was reversed-transcribed in three overlapping cDNA segments (RT-PCR) which were amplified and further analyzed by PTT after in vitro transcription/translation.


RESULTS
This strategy enabled us to detect a truncated peptide in each of the 6 unrelated patients from southern France who were investigated. The mutation was further characterized by direct sequencing of the RT-PCR product.


CONCLUSIONS
In CHM gene, all conditions are present to make the RT-PCR-PTT strategy the method of choice for mutation screening. As a result of the simplified protocol described in this study, the families of the patients could benefit from accurate carrier-status assessment.
Laboratoire de Biochimie Génétique, CNRS ERS-155, Institut de Biologie, Montpellier, Francè.
L
L
Beaufrere
A
A
Girardet
B
B
Arnaud
M
M
Claustres
S
S
Tuffery
D048868:Adaptor Proteins, Signal Transducing; D019883:Alkyl and Aryl Transferases; D002352:Carrier Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D015794:Choroideremia / Q000175:diagnosis* / Q000235:genetics; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000235:genetics; D005136:Eye Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D005260:Female; D005602:France; D005808:Genes, Recessive; D006580:Genetic Carrier Screening / Q000379:methods*; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D008297:Male; D014176:Protein Biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014960:X Chromosome / Q000235:genetics*; D020691:rab GTP-Binding Proteins
D048868:Adaptor Proteins, Signal Transducing; C081787:CHM protein, human; D002352:Carrier Proteins; D018076:DNA, Complementary; D005136:Eye Proteins; D019883:Alkyl and Aryl Transferases; D020691:rab GTP-Binding Proteins
Mise au point d'un test diagnostique de la choroïdérémie: le test de troncation des protéines (PTT).