title: New molecular beacon for p53 gene point mutation and significant potential in serving as the polymerization primer.
abstract: Molecular beacon (MB) is usually explored as a convenient probe for various bioassays. In an enzymatic polymerization-based biosensing system, primer, and MB, sometimes involving other oligonucleotides, are often required to collaboratively generate an amplified fluorescent signal to detect target molecules with high sensitivity and specificity. In the current study, a multifunctional primer-integrated MB (MP-MB) was developed to detect the p53 tumor suppressor gene. Compared with the traditional MB, our MP-MB can not only selectively identify the target of interest and signal sensitively its hybridization event, but also act as the primer during enzymatic polymerization. Specifically, hybridization of MP-MB to target p53 gene restored the fluorescence intensity and activated the pre-locked primer designed by changing the molecular configuration of MP-MB. Moreover, the p53 gene could be detected down to 1nM with a linear response range of 1×10(-9)-3×10(-7)M, and p53 gene point mutation was readily distinguished from the wild-type one. Its potential application as a primer of replication in enzymatic polymerization-based assay systems was validated by running parallel gel electrophoreses in comparison with the native counterpart of MP-MB without any chemical modification. Owning to its excellent assay characteristics, less species requirement, broad sequence diversity and preserved intrinsic bioactivity, the proof-of-concept of MP-MB exhibits a great potential in various biomedical applications.
authors.affiliation: Cancer Metastasis Alert and Prevention Center, Pharmaceutical Photocatalysis, State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Xu
authors.affiliation: Cancer Metastasis Alert and Prevention Center, Pharmaceutical Photocatalysis, State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Dong
authors.affiliation: Cancer Metastasis Alert and Prevention Center, Pharmaceutical Photocatalysis, State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China.
authors.forename: Weiyu
authors.initials: W
authors.lastname: Shen
authors.affiliation: Cancer Metastasis Alert and Prevention Center, Pharmaceutical Photocatalysis, State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China.
authors.forename: Sudan
authors.initials: S
authors.lastname: He
authors.affiliation: Cancer Metastasis Alert and Prevention Center, Pharmaceutical Photocatalysis, State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China.
authors.forename: Hongling
authors.initials: H
authors.lastname: Li
authors.affiliation: Cancer Metastasis Alert and Prevention Center, Pharmaceutical Photocatalysis, State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China.
authors.forename: Yusheng
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Cancer Metastasis Alert and Prevention Center, Pharmaceutical Photocatalysis, State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China; State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, China. Electronic address: wuzaisheng@163.com.
authors.forename: Zai-Sheng
authors.initials: ZS
authors.lastname: Wu
authors.affiliation: Cancer Metastasis Alert and Prevention Center, Pharmaceutical Photocatalysis, State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China. Electronic address: pharmlink@gmail.com.
authors.forename: Lee
authors.initials: L
authors.lastname: Jia
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics*; D005456:Fluorescent Dyes; D016158:Genes, p53; D006801:Humans; D025202:Molecular Diagnostic Techniques; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000235:genetics*; D017354:Point Mutation; D058105:Polymerization
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes
keywords: Multifunctional primer-integrated MB (MP-MB); Polymerization amplification; p53 point mutation