scholarly journals Detecting familial defective apolipoprotein B-100: three molecular scanning methods compared

1997 ◽  
Vol 43 (9) ◽  
pp. 1630-1634 ◽  
Author(s):  
Bryony G Henderson ◽  
Philip R Wenham ◽  
J Peter Ashby ◽  
Gillian Blundell
Keyword(s):  
Apolipoprotein B ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Single Strand Conformation Polymorphism ◽  
Sscp Analysis ◽  
Heteroduplex Analysis ◽  
Apo B ◽  
Large Numbers ◽  
Gradient Gel Electrophoresis ◽  
Conformation Polymorphism

Abstract Familial defective apolipoprotein (apo) B-100 (FDB), a condition that may give rise to hypercholesterolemia, is caused by mutations around codon 3500 of the apo B gene. We have compared the ability of three molecular-scanning techniques, heteroduplex analysis, single-strand conformation polymorphism (SSCP) analysis, and denaturing gradient gel electrophoresis (DGGE), to detect these mutations in a cohort of 432 hypercholesterolemic individuals. Heteroduplex analysis and DGGE detected 11 individuals with apo B mutations, 9 of whom were heterozygous for apo B R3500Q and 2 who were heterozygous for apo B R3531C. Whereas DGGE was able to distinguish between these two mutations, heteroduplex analysis was technically simpler and gave a higher sample throughput. In contrast, SSCP analysis detected only 7 of the R3500Q and none of the R3531C heterozygotes and was the most complex of the three techniques. We believe heteroduplex analysis to be the method of choice for screening large numbers of samples for FDB.

Mutation screening of the codon 3500 region of the apolipoprotein B gene by denaturing gradient-gel electrophoresis

1995 ◽  
Vol 41 (3) ◽  
pp. 419-423 ◽  
Author(s):  
H Nissen ◽  
P S Hansen ◽  
O Faergeman ◽  
M Hørder
Keyword(s):  
Apolipoprotein B ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Low Density Lipoprotein ◽  
Mutation Screening ◽  
Screening Method ◽  
Density Lipoprotein ◽  
Apo B ◽  
Density Lipoprotein Receptor ◽  
Gradient Gel Electrophoresis

Abstract Familial defective apolipoprotein B (FDB) is a clinical condition resembling familial hypercholesterolemia. The underlying genetic defects are mutations in the apolipoprotein B-100 (apo B-100) gene. Two mutations (Arg3500 --> Gln and Arg3531 --> Cys) are known to date. We designed a denaturing gradient-gel electrophoresis (DGGE) technique to detect sequence variations in codons 3456-3553 of the apo B-100 gene. In 46 heterozygous FDB patients with the predominant codon 3500 mutation, a uniform four-band DGGE pattern was seen, whereas 57 non-FDB patients showed the uniform single-band pattern expected in normal homozygotes. The recently described codon 3531 mutation and a previously unpublished Arg --> Pro mutation in codon 3480 showed unique DGGE patterns, allowing unambiguous differentiation of the three mutations. The DGGE method thus both detects known FDB mutations and screens for other mutations in codons 3456-3553 of the low-density lipoprotein receptor binding region of apo B-100; it can be used as a first-line screening method for FDB.

scholarly journals Identifikasi Biodeteriogen Sebagai Langkah Awal Konservasi Benda Cagar Budaya

2016 ◽  
Vol 10 (2) ◽  
pp. 23-30
Author(s):  
Moh Habibi
Keyword(s):  
Temperature Gradient ◽  
Clone Library ◽  
Ribosomal Dna ◽  
Gel Electrophoresis ◽  
Restriction Analysis ◽  
Single Strand Conformation Polymorphism ◽  
Gradient Gel Electrophoresis ◽  
Dna Restriction ◽  
Temperature Gradient Gel Electrophoresis ◽  
Conformation Polymorphism

Indonesia mempunyai Benda Cagar Budaya (BCB) yang berlimpah. BCB tersebut dapat mengalami degradasi disebabkan mikroorganisme. Penelitian ini bertujuan untuk memberikan pengetahuan mengenai teknik molecular yang digunakan untuk deteksi biodeteriogen pada benda cagar budaya. Teknik Molekuler yang dapat digunakan adalah Fingerprinting, meliputi DGGE (Denaturing Temperature Gradient Gel Electrophoresis), SSCP (Single Strand Conformation Polymorphism), ARDRA (Amplified Ribosomal DNA Restriction Analysis), dan Clone Library. Teknik ini mempunyai beberapa kelebihan, seperti tingkat presisi yang tinggi, lebih cepat digunakan, dapat digunakan untuk mengetahui peran biodeteriogen pada lingkungan.

scholarly journals Denaturing gradient-gel electrophoresis screening of familial defective apolipoprotein B-100 in a mixed Asian cohort: two cases of arginine3500 → tryptophan mutation associated with a unique haplotype

1997 ◽  
Vol 43 (6) ◽  
pp. 916-923 ◽  
Author(s):  
Meng-Ling Choong ◽  
Evelyn S C Koay ◽  
Kah-Lin Khoo ◽  
Min-Cheh Khaw ◽  
Sunil K Sethi
Keyword(s):  
Receptor Binding ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Ldl Receptor ◽  
Pcr Amplification ◽  
Unique Haplotype ◽  
Apo B ◽  
Putative Receptor ◽  
Published Report ◽  
Gradient Gel Electrophoresis

Abstract The Arg-to-Trp substitution at codon 3500 in the apolipoprotein (apo) B-100 gene is established as a cause of familial defective apo B-100 (FDB), a functional mutation, resulting in reduced LDL receptor binding and manifest hypercholesterolemia. In a search for similar mutations in 163 Malaysians, we screened the putative receptor-binding region (codons 3456–3553) of the apo B-100 gene by PCR amplification and denaturing gradient-gel electrophoresis. Four single-base mutations were detected and confirmed by DNA sequencing. Two females, a Chinese and a Malay, had the same CGG3500 → TGG mutation, resulting in an Arg3500-to-Trp substitution. This is the second published report of such an independent mutation involving the same codon as the established Arg3500-to-Gln mutation. The two other mutations detected, CTT3517 → CTG and GCC3527 → GCT, resulted in degenerate codons with no amino acid substitutions. All four mutations were associated with a unique apo B haplotype, different from those found in Caucasian FDB patients but concurring with that previously reported for two other Asians with FDB.

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