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Original ArticleOpen Access
Screening for Mutations in Exons Encoding the Ligand-Binding Domain of the LDL Re~eptor Gene Using PCR-CFLP and PCR-SSCP
Pongrapeepom K 
,
Yamwong P ,
Leowattana W ,
OngAjyooth S ,
Nuchpramool W ,
Amomrattana A ,
Kerdsaeng K ,
Kasemsuk L ,
Thepsuriyanon P ,
Laungsuwan S ,
Kiartivich S ,
Sribhen K
,
Yamwong P ,
Leowattana W ,
OngAjyooth S ,
Nuchpramool W ,
Amomrattana A ,
Kerdsaeng K ,
Kasemsuk L ,
Thepsuriyanon P ,
Laungsuwan S ,
Kiartivich S ,
Sribhen K PREYANUJ YAMWONG, M.D.**,
SOMPONG ONG-AJYOOTH, M.Sc. *,
ANCHALEE AMORNRATTANA, M.Sc. **,
LUMPOON KASEMSUK, B.Sc.*,
SIV ADEE LAUNGSUW AN, M.Sc. ***,
KOSIT SRIBHEN, M.D.***
Primary hypercholesterolemia includes both monogenic disorders and polygenic condi-
tions. Two well defined monogenic disorders are familial hypercholesterolemia (FH) and familial
defective apolipoprotein (apo) B-1 00 (FDB). Both disorders convey high risk of premature coro-
nary artery disease. FH and FDB are caused by mutations in LDL receptor and apo B-1 00 genes,
respectively. In the present study, mutations in both genes in Thai subjects with primary hyper-
cholesterolemia were screened. For apo B-100 gene, a common mutation R3500Q was screened.
This mutation was not observed in the patients (n
=
45). For LDL receptor gene, mutations in the
exons encoding the ligand -binding domain were screened. By PCR-CFLP analysis, 18 abnormal
CFLP patterns in exon 4, the hot spot for mutations, were found in patients (n=45). One of the
DNA samples with abnormal CFLP patterns was previously identified and reported as a possible
disease-causing mutation, namely Dl51Y. For the other exons, the screening technique was PCR-
SSCP. Abnormal SSCP patterns in DNA samples from patients (n=20) were found as follows, two
in exon 3, one in exon 5 and another one in exon 6. Further characterization by DNA sequencing
and family studies for these abnormal patterns are underway.
Key word : Hypercholesterolemia, LDL Receptor Gene, Mutation. PCR-CFLP, PCR-SSCP
SOMPONG ONG-AJYOOTH, M.Sc. *,
ANCHALEE AMORNRATTANA, M.Sc. **,
LUMPOON KASEMSUK, B.Sc.*,
SIV ADEE LAUNGSUW AN, M.Sc. ***,
KOSIT SRIBHEN, M.D.***
Primary hypercholesterolemia includes both monogenic disorders and polygenic condi-
tions. Two well defined monogenic disorders are familial hypercholesterolemia (FH) and familial
defective apolipoprotein (apo) B-1 00 (FDB). Both disorders convey high risk of premature coro-
nary artery disease. FH and FDB are caused by mutations in LDL receptor and apo B-1 00 genes,
respectively. In the present study, mutations in both genes in Thai subjects with primary hyper-
cholesterolemia were screened. For apo B-100 gene, a common mutation R3500Q was screened.
This mutation was not observed in the patients (n
=
45). For LDL receptor gene, mutations in the
exons encoding the ligand -binding domain were screened. By PCR-CFLP analysis, 18 abnormal
CFLP patterns in exon 4, the hot spot for mutations, were found in patients (n=45). One of the
DNA samples with abnormal CFLP patterns was previously identified and reported as a possible
disease-causing mutation, namely Dl51Y. For the other exons, the screening technique was PCR-
SSCP. Abnormal SSCP patterns in DNA samples from patients (n=20) were found as follows, two
in exon 3, one in exon 5 and another one in exon 6. Further characterization by DNA sequencing
and family studies for these abnormal patterns are underway.
Key word : Hypercholesterolemia, LDL Receptor Gene, Mutation. PCR-CFLP, PCR-SSCP
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