Advancements in Characterization Techniques of Biopolymers: Cyclic Voltammetry, Gel Electrophoresis, Circular Dichroism and Fluorescence Spectroscopy

Kaushik, Mahima; Kumar, Manisha

doi:10.4172/2379-1764.1000184

Cited by 1 publication

(4 citation statements)

References 76 publications

(78 reference statements)

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“…Figure a shows the CD spectra of APOE22T and APOE22C at 3 μM strand concentration in 20 mM sodium cacodylate buffer (pH 7.4) comprising 100 mM NaCl and 0.1 mM EDTA. Both APOE22T and APOE22C displayed a strong positive peak at 287 nm and a strong negative peak at 249 nm, which are the characteristic CD signatures of B-form of DNA. , …”

Section: Resultsmentioning

confidence: 95%

“…The molecularity of the DNA structures can be established by carrying out native polyacrylamide gel electrophoresis. 13 A 15% gel electrophoretogram having the DNA samples of APOE22T as well as APOE22C along with their duplexes was run in 20 mM sodium cacodylate buffer (pH 7.4) containing 100 mM NaCl and 0.1 mM ethylenediaminetetraacetic acid (EDTA, Figure 2 a). 12-mer and 24-mer duplex (PAL12 and PAL24) as well as 35-mer single-stranded random sequence (M35) were used as control size markers.…”

Section: Resultsmentioning

confidence: 99%

“…Both APOE22T and APOE22C displayed a strong positive peak at 287 nm and a strong negative peak at 249 nm, which are the characteristic CD signatures of B-form of DNA. 13 , 14 …”

Section: Resultsmentioning

confidence: 99%

“…Both APOE22T and APOE22C displayed a strong positive peak at 287 nm and a strong negative peak at 249 nm, which are the characteristic CD signatures of B-form of DNA. 13,14 Oligomer Concentration Dependence in CD Studies. Because the results obtained from oligomer dependence in gel electrophoresis experiments suggested the formation of a new structure at a high oligomer concentration in case of APOE22T, further CD experiments were performed to get more clarity on the same.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Structural Switch from Hairpin to Duplex/Antiparallel G-Quadruplex at Single-Nucleotide Polymorphism (SNP) Site of Human Apolipoprotein E (APOE) Gene Coding Region

Chaudhary¹,

Kaushik²,

Ahmed³

et al. 2018

ACS Omega

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A gradual dementia, which leads to the loss of memory and intellectual abilities, is the main characteristics of Alzheimer’s disease. Amyloid-β (Aβ) plaques are the main components that accumulate and form clumps in the brains of people suffering from Alzheimer’s disease. Apolipoprotein E (APOE), an amyloid-binding protein is considered as one of the main genetic risk factor of the late-onset Alzheimer’s disease. Different isoforms of APOE gene named APOE2, APOE3, and APOE4 are known to exist, which differ from each other at certain positions involving single-nucleotide polymorphisms (SNPs). Out of these isoforms, APOE4 increases the risk of developing late-onset Alzheimer’s disease, whereas APOE3 is the most common among the general population. APOE4 differs from the common APOE3 by only one nucleotide at position +2985 (T to C), which results in immense alteration in the structure and function of the APOE gene. A combination of gel electrophoresis (polyacrylamide gel electrophoresis, PAGE), circular dichroism (CD), CD melting, thermal difference spectra and UV-thermal denaturation (TM) techniques was used to investigate the structural polymorphism associated with T → C single-nucleotide polymorphism (SNP) at the GC-rich sequence (d-TGGAGGACGTGTGCGGCCGCCT; APOE22T). Herein, we report that APOE22T DNA sequence switches between hairpin to antiparallel quadruplex from low to high oligomer concentration. On the contrary, its C-counterpart (APOE22C) forms hairpin as well as intermolecular antiparallel duplex structure. This structural change may possibly contribute to the protein recognition pattern, which, in turn, might control the APOE gene expression.

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Section: Resultsmentioning

confidence: 95%

Section: Resultsmentioning

confidence: 99%

“…Both APOE22T and APOE22C displayed a strong positive peak at 287 nm and a strong negative peak at 249 nm, which are the characteristic CD signatures of B-form of DNA. 13 , 14 …”

Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

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