Citation: LIU Hui-Min, HAO Ju-Min, XU Jing, YAN Qing-Pi, GAO Han, ZHENG Jiang. Selection and Identification of Common Aptamers Against Both Vibrio Harveyi and Vibrio Alginolyticus[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(5): 623-631. doi: 10.19756/j.issn.0253-3820.191663 shu

Selection and Identification of Common Aptamers Against Both Vibrio Harveyi and Vibrio Alginolyticus

LIU Hui-Min ^1,2, ,
HAO Ju-Min ¹ ,
XU Jing ¹ ,
YAN Qing-Pi ¹ ,
GAO Han ¹ ,
ZHENG Jiang ¹

1.
Engineering Research Center of the Modern Technology for Eel Industry, Ministry of Education, Engineering Research Center of Aquaculture Breeding and Healthy of Fujian, Fisheries College of Jimei University, Xiamen 361021, China;
2.
Fujian Province Key Laboratory of Special Aquatic Formula Feed, Fujian Tianma Science and Technology Group Co. Ltd., Fuqing 350308, China

Received Date: 11 November 2019
Revised Date: 16 December 2019

Fund Project: This work was supported by the Natural Science Foundation of Fujian Province, China (No. 2018J01455), the Open Fund of Engineering Research Center of the Modern Technology for Eel Industry of Ministry of Education of China (No. RE201808), the Open Research Fund Program of Fujian Engineering Research Center of Aquatic Breeding and Healthy Aquaculture (No. DF201901), and the Open Research Fund from Fujian Province Key Laboratory of Special Aquatic Formula Feed (Fujian Tianma Science and Technology Group Co., Ltd.) (No. TMKJZ1909).

Common sites are the sites of same or similar structure on different microorganisms. Some of the common sites can be shared by same genus of microorganisms, which can be used as the marker for the identification of the microorganism. Therefore, selection of common aptamers against these common sites is not only helpful to the analysis of the microorganisms, but also of great significance for the identification of the relative microorganisms. In the work, the common aptamers against both Vibiro harveyi and Vibiro alginolyticus were selected after 8 rounds of selection. The affinity of the enrichment pool of the aptamers increased by 24.1 times, and the analysis based on homologous tree showed that the sequences of the aptamer enriched pool showed obvious convergence, which showed a good selection efficiency. Two aptamers (C14 and C22) were proved for good affinity and specificity towards both V. harveyi and V. alginolyticus. The affinities between the two aptamers and V. harveyi were significantly stronger than those between the aptamers and V. alginolyticus. The affinity constants (K _d) of the aptamer C14 against V. harveyi and V. alginolyticus were 55.76 and 82.88 nmol/L respectively. And the affinity constants of the aptamer C22 against V. harveyi and V. alginolyticus were 33.97 and 43.95 nmol/L respectively. Secondary structures of the two aptamers were mainly composed of stems and loops. The 3'-end structure of aptamer C22 seemed to be more complex than that of aptamer C14. Significant fluorescence could be found only on V. harveyi and V. alginolyticus after the aptamers marked with fluorescence group were mixed with V. harveyi, V. alginolyticus, Aeromonas hydrophila, Edwardsiella tarda and Escherichia coli, which showed that the two aptamers had good affinity and specificity towards the two target bacteria. This studies laid a foundation for the identification and detection of the two microorganisms as well as the aquatic pathogenic vibrios.
- Vibrio harveyi,
- Vibrio alginolyticus,
- Common aptamer,
- Selection,
- Fluorescence microscope
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