scholarly journals Fluorescence-based, Nonradioactive Method for Efficient Detection of the Pentanucleotide Repeat (TTTTA)n Polymorphism in the Apolipoprotein(a) Gene

2001 ◽  
Vol 47 (10) ◽  
pp. 1758-1762 ◽  
Author(s):  
Jill Rubin ◽  
Thomas A Pearson ◽  
Roberta G Reed ◽  
Lars Berglund
Keyword(s):  
African Americans ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Reading Frame ◽  
Apolipoprotein A ◽  
Efficient Detection ◽  
Pcr Products ◽  
Major Predictor ◽  
Polymorphism Pattern ◽  
Pentanucleotide Repeat

Abstract Background: The apolipoprotein(a) [apo(a)] gene is a major predictor of plasma lipoprotein(a) concentrations, an independent risk factor for cardiovascular disease. The apo(a) gene contains a pentanucleotide repeat (PNR) polymorphism, 1.4 kb upstream from the apo(a) gene reading frame. This polymorphism has been suggested to be important in control of apo(a) gene expression. Methods: We developed a fluorescence-based, nonradioactive procedure to detect the PNR polymorphism. After amplification of the polymorphism by PCR, the respective PCR products were separated by denaturing polyacrylamide gel electrophoresis and detected using a 3′-end fluorescently labeled oligonucleotide as a probe. We used the method to characterize the PNR polymorphism pattern in 313 individuals, 195 Caucasians and 118 African Americans. The new method efficiently separated DNAs corresponding to the different PNR repeats. Results: Among both ethnic groups, alleles containing eight PNRs were most common. Smaller PNRs were more common among African Americans, and larger PNRs were more common among Caucasians. Conclusions: We developed a nonradioactive technique that separates the PNR polymorphism in the apo(a) gene and can be used in other studies involving closely sized polymorphisms.

Identification of Apo(a) Phenotypes in a Korean Population Using a Standardized Nomenclature System Based on the Number of Kringle IV Repeats

1997 ◽  
Vol 34 (6) ◽  
pp. 681-687
Author(s):  
Won-Ki Min ◽  
Jae Ok Lee ◽  
Chun Hee Kim ◽  
Junghan Song ◽  
Jin Q Kim
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Serum Lipoprotein ◽  
Allele Size ◽  
Nomenclature System ◽  
Molecular Weights ◽  
Apolipoprotein A ◽  
Electrophoresis Method

We determined serum lipoprotein(a) [Lp(a)] concentrations and apolipoprotein(a) [apo(a)] phenotypes in 193 healthy Koreans. We analysed the apo(a) phenotypes by a simplified sodium dodecyl sulphate-polyacrylamide gel electrophoresis method and classified apo(a) isoforms objectively using an apo(a) phenotype standard with a known number of kringle IV repeats. The frequency distribution of Lp(a) levels showed marked positive skew with a mean of 0·143 g/L and a median of 0·052 g/L. Of the 193 subjects tested, no bands were detected in three, and single- and double-band phenotypes were observed in 103 and 87, respectively. Among the Koreans, the most frequent phenotype was S5(39·4%), followed by S4S5(17·1%), S5S5(14·0%), S4(11·4%), S3S5(5·2%), and the remaining phenotypes (13·0%). The calculated apo(a) allele frequencies were LpF, 0·003; LpS1, 0·013; LpS2, 0·010; LpS3 0·048; LpS4 0·198; LpS5, 0·563 and Lp0, 0·165. We found that the serum Lp(a) concentration in Koreans was similar to that of Caucasians, but the apo(a) allele size distribution was shifted toward higher molecular weights.

scholarly journals An Insertion/Deletion Polymorphism within the Promoter of EGLN2 is Associated with Susceptibility to Colorectal Cancer

2017 ◽  
Vol 32 (3) ◽  
pp. 274-277 ◽  
Author(s):  
Chaoyang Li ◽  
Lanjun Feng ◽  
Luwei Niu ◽  
Teng Teng Li ◽  
Bin Zhang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Gel Electrophoresis ◽  
Chinese Population ◽  
Functional Role ◽  
Polyacrylamide Gel Electrophoresis ◽  
Proximal Promoter ◽  
Deletion Polymorphism ◽  
Increased Risk ◽  
Codominant Model ◽  
Pcr Products

Objective To investigate the association between susceptibility to colorectal cancer (CRC) and a 4-bp insertion/deletion polymorphism (rs10680577) in the proximal promoter of the EGLN2 gene. Method The first step in genotyping EGLN2 was PCR, then the PCR products were separated using 7% nondenaturing polyacrylamide gel electrophoresis and visualized by silver staining according to the final product band location and quantity to determine the genotype of the sample. The final count was done by two different pathologists. Result In the codominant model, compared with the ins/ins genotype, subjects with the heterozygous ins/del or homozygous del/del genotype had a significantly increased risk of CRC (adjusted OR = 1.45, p<0.0001 and OR = 2.44, p = 0.0001, respectively). Each additional copy of the 4-bp deletion allele conferred a significantly increased risk of CRC (OR = 1.47, 95% CI 1.28-1.66, p<0.0001). In the stratification analysis, we further proved that the association was more prominent in TNM stage III and IV cancer compared with stage I and II (adjusted OR = 1.43, 95% CI 1.07-1.93, p for heterogeneity = 0.02). Conclusions Our study provided initial evidence that the insertion/deletion polymorphism rs10680577 may play a functional role in the development of CRC in the Chinese population.

Heterogeneity of lipoprotein Lp(a) and apolipoprotein(a).

1988 ◽  
Vol 34 (6) ◽  
pp. 1036-1040 ◽  
Author(s):  
G F Grinstead ◽  
R D Ellefson
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Core Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Single Patient ◽  
Selective Precipitation ◽  
Apolipoprotein A ◽  
Molecular Masses ◽  
Chemical Deglycosylation

Abstract We have purified Lp(a) lipoproteins from sera of four subjects by ultracentrifugation, selective precipitation, and chromatofocusing. Each subject had two forms of serum Lp(a) that were separable by chromatofocusing. We purified apolipoprotein (a) [apo(a)] from the eight isolated Lp(a)s and obtained only one form of apo(a) from each subject. The four apo(a)s seen on sodium dodecyl sulfate-polyacrylamide gel electrophoresis had different apparent molecular masses, ranging from 275 to 440 kDa. Chemical deglycosylation of the smallest apo(a) yielded a 235 kDa protein, which may be a core protein structure common to all apo(a)s. We conclude that there are many forms of serum Lp(a) and apo(a). The heterogeneity of serum Lp(a) particles can be ascribed in part to differences in size of apo(a), but other factors must account for the existence within a single patient of different Lp(a)s that contain apparently identical apo(a). One must consider the heterogeneity of Lp(a) when designing assays for this lipoprotein.

Rapid microsatellite analysis using discontinuous polyacrylamide gel electrophoresis

Genome ◽  
2002 ◽  
Vol 45 (6) ◽  
pp. 1107-1109 ◽  
Author(s):  
E White ◽  
R Sahota ◽  
S Edes
Keyword(s):  
High Resolution ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Population Data ◽  
Microsatellite Analysis ◽  
Disc Electrophoresis ◽  
Polyacrylamide Gel ◽  
Large Numbers ◽  
Pcr Products ◽  
Slab Gels

A method for screening large numbers of samples for microsatellites using discontinuous, non-denaturing polyacrylamide gels and rapid fluorescent gel staining is described. Disc electrophoresis on slab gels provides high-resolution of PCR products. It is useful for collecting population data once microsatellite loci have been characterized.Key words: microsatellite, discontinuous polyacrylamide gel electrophoresis, non-denaturing

Analysis for Cerebrospinal Fluid Proteins by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis

1992 ◽  
Vol 38 (10) ◽  
pp. 2008-2012 ◽  
Author(s):  
F Mashige ◽  
T Shimizu ◽  
S Iijima ◽  
A Ohkubo
Keyword(s):  
Cerebrospinal Fluid ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Apolipoprotein A ◽  
Csf Proteins ◽  
Sds Page ◽  
Molecular Masses

Abstract Cerebrospinal fluid (CSF) proteins with molecular masses of &lt; 150,000 Da were identified by immunoblotting after two kinds of nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). With PAGE 1 (17-27% gradient gel), CSF proteins were clearly separated into seven to nine bands with molecular masses of 3000-67,000 Da; seven bands were identified as beta 2-microglobulin, lysozyme, prealbumin, free kappa and lambda chain, apolipoprotein A-I, glycoproteins, and albumin by immunoblotting. With PAGE 2 (10-20% gradient gel), proteins were clearly separated into 11-16 bands with molecular masses of 15,000-150,000 Da; 11 were identified as prealbumin, free kappa and lambda chain, apolipoprotein A-I, glycoproteins, albumin, alpha 1-antitrypsin, transferrin (separated into two bands), immunoglobulin fragments, haptoglobin, and IgG. We analyzed CSF samples collected from 81 patients with cerebrospinal signs by these SDS-PAGE methods and observed prominent bands in some cases.

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