Combination and cleavage of HBV DNA fragments by triple helix-forming oligonucleotides modified with manganese porphyrin in vitro.
- Author:
Lixia GUANG
1
;
Fahuan YUAN
;
Min XI
;
Congmin ZHAO
;
Li LIU
;
Enyi WEN
;
Youping AI
Author Information
- Publication Type:Journal Article
- MeSH: DNA; drug effects; pharmacology; DNA, Viral; chemistry; drug effects; Hepatitis B virus; genetics; Manganese; pharmacology; Metalloporphyrins; pharmacology; Potassium Compounds; pharmacology; Sulfates; pharmacology
- From: Chinese Medical Journal 2003;116(8):1248-1252
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo observe the ability of triple helix-forming oligonucleotides (TFOs) modified with manganese porphyrin to combine with and cleave HBV DNA fractions.
METHODSTFO were modified with manganese porphyrin and acridines, and then reacted with the (32)P labeled HBV DNA fragments at 37 degrees C in vitro (pH 7.4). Electrophoretic mobility shift assays and DNase I footprinting tests were used to show the affinity and specificity of TFO to bind to target sequences. The ability of TFO to cleave HBV DNA fragments was tested by cleavage experiments.
RESULTSTFO modified with manganese porphyrin and acridine could bind to the target sequence in a sequence-dependent manner, with a Kd value of 3.5 x 10(-7) mol/L and a relative affinity of 0.008. In the presence of potassium monopersulfate (KHSO(5)), TFO modified with manganese porphyrin and acridine could cleave the target sequence where the triplex DNA was formed.
CONCLUSIONIn the presence of KHSO(5), TFO modified with manganese porphyrin and acridine could bind and cleave the target HBV-DNA in a sequence-dependent manner.
