Paper Title:
In Vitro Selection of Oligonucleotide Acid Aptamers against Pathogenic Vibrio Alginolyticus by SELEX
  Abstract

Detection of pathogenic microorganism is very necessary in the control of infectious disease prevailing in aquiculture animals. However, most of the present techniques can not meet the need of the quick field inspection. Systematic evolution of ligands by exponential enrichment (SELEX) is a new molecular recognition way for generating high affinity oligonucleotide acid aptamers, a new nucleotide acid material, which have been widely used in the detections of proteins, cells and so on. In the present paper, the technology was applied to select the high affinity aptamers against pathogenic microorganism Vibrio alginolyticus, which could be used for the rapid field detection of the microorganism. Based on the designment of the ssDNA library of 76 nucleotide acids with 35-base random region, the SELEX system for the selection of the high affinity aptamers against Vibrio alginolyticus was established. In the SELEX system, asymmetric PCR was proved to be a better amplification method for the ssDNA library than the reported affinity magnetic bead method, and the corresponding parameters of the asymmetric PCR were also studied and optimized. The affinity of the final ssDNA library increased by nearly 200% compared with the original library. Cloning and sequencing of the final ssDNA library showed that there were at least two kinds of ssDNAs with different length in the affinity ssDNA library: one was 76 bases, another was 149 bases. Simulation of the secondary structures showed that the secondary structures of the two fragments were different greatly, suggesting that the two fragments could bind to different sites of V. alginolyticus surface.

  Info
Periodical
Key Engineering Materials (Volumes 439-440)
Edited by
Yanwen Wu
Pages
1456-1462
DOI
10.4028/www.scientific.net/KEM.439-440.1456
Citation
J. Zheng, Y. B. Li, J. X. Li, J. Wang, Y. Q. Su, "In Vitro Selection of Oligonucleotide Acid Aptamers against Pathogenic Vibrio Alginolyticus by SELEX", Key Engineering Materials, Vols. 439-440, pp. 1456-1462, 2010
Online since
June 2010
Export
Price
$32.00
Share

In order to see related information, you need to Login.

In order to see related information, you need to Login.

Authors: You Rui Huang, Hai Bo Hu
Abstract:PID control scheme has been widely used in most of control system. The method of design PID controller is mature gradually. Due to the...
1061
  | Authors: Fang Zhu, Jun Fang Wei
Chapter 3: Hardware, Information Technology and System
Abstract:The performance of support vector machine (SVM) depends on the selection of model parameters, however, the selection of SVM model parameters...
694
Authors: Sofiah Hamzah, Nora'aini Ali, Marinah Mohd Ariffin, Abdul Wahab Mohammad
Chapter 4: Separation Processes and Membranes, Thermodynamics
Abstract:This study aimed to characterize a highly specific affinity membrane for trypsin separation. The basic membrane was fabricated using 15%...
592
Authors: Rui Zhao, Yong Zheng Wen, Wei Ma, Jian Cheng Yang, Xiao Mei Yu
Chapter 7: Bioresearch and Biomedical Engineering
Abstract:In this paper, an aptasensor with high sensitivity and rapid response was developed for the detection of staphylococcus enterotoxin B (SEB)...
1298
Authors: Guo Yin Huang, Long Fei Ma, Ming Yuan Guan, Jin Tao Liang, Yong Huang, Gui Yin Li
Chapter 2: Nanotechnologies and Nanomaterials: Frontiers of Application and Properties
Abstract:In this paper, a novel hIg E aptamer biosensor was designed base on the core-shell Fe3O4@Au magnetic composite...
170