In vitro expression study of novel mutations in phenylalanine hydroxylase gene / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 673-677, 2013.
Article
in Chinese
| WPRIM
| ID: wpr-254538
ABSTRACT
<p><b>OBJECTIVE</b>To study the in vitro expression of 6 novel missense mutations (R270G, P275A, F121L, A156P, E183G, I324N) and a previously described R408Q mutation of phenylalanine hydroxylase (PAH) gene and explore the genotype-phenotype correlation through comparison of protein levels and residual enzyme activities.</p><p><b>METHODS</b>Seven expression vectors containing PAH cDNA were constructed with a site-directed mutagenesis kit. The plasmids were extracted and sequenced to confirm the target mutations. pcDNA3.0 containing PAH cDNA was transfected into COS-7 cells and total proteins were extracted 48 h after transfection. The quantities of proteins and residual enzyme activities of the 7 mutants were assessed with the wild-type PAH gene as reference.</p><p><b>RESULTS</b>Relative quantities of PAH proteins for R270G, P275A, F121L, A156P, E183G, I324N and R408Q were 10.5%, 56.6%, 54.3%, 8.7%, 8.5%, 67.3% and 85.4%, respectively. The residual enzyme activities were 7.7%, 27.6%, 19.0%, 10.4%, 9.1%, 50.6% and 40.2%, respectively.</p><p><b>CONCLUSION</b>PAH residual enzyme activities of 7 PAH mutants were all significantly reduced.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Phenylalanine Hydroxylase
/
Molecular Sequence Data
/
Chlorocebus aethiops
/
Sequence Alignment
/
Amino Acid Sequence
/
COS Cells
/
Mutation, Missense
/
Genetic Association Studies
/
Genetics
/
Methods
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Medical Genetics
Year:
2013
Type:
Article
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