适于检测非洲猪瘟病毒的点亮Spinach-p54 RNA适配体的设计及应用

doi:10.16288/j.yczz.21-335

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Hereditas(Beijing) ›› 2021, Vol. 43 ›› Issue (12): 1170-1178.doi: 10.16288/j.yczz.21-335

• Technique and Method • Previous Articles Next Articles

Design and application of lightening Spinach-p54 RNA apatmer to detect African swine fever virus

Chengcheng Han^1,²(), Kai Xia^1,³, Ruying Gong¹, Xuhan Wu¹, Lei Zhang^1,²(), Xinle Liang^1,²()

1. School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310018, China
2. Institute of Food Biotechnology, Zhejiang Gongshang University, Hangzhou 310018, China
3. School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou 310023, China

Received:2021-09-21 Revised:2021-11-30 Online:2021-12-20 Published:2021-12-10
Contact: Zhang Lei,Liang Xinle E-mail:707884766@qq.com;zhanglei@zjsu.edu.cn;dbiot@mail.zjgsu.edu.cn

Abstract

Abstract:

African swine fever virus (ASFV) is a major and prevalent pathogenic virus in livestock. Its outbreak in the recent years has seriously affected the national economic life. In this study, a specific chimeric RNA aptamer Spinach-p54 corresponding to the conserved protein p54 of ASFV, was designed and used in assembling a toehold RNA module with a truncated lightening Spinach switch. They were used to detect the p54 gene. The aptamer bound rapidly and paired specifically with the ASFV p54 target sequence and produced fluorescence with the addition of DFHBI substrate. The results showed that the limit of detection was as low as 200 nmol/L p54 fragment, under the conditions of 200 nmol/L Spinach-p54 aptamer and 37 ℃, suggesting that the current method can provide a simple, specific, sensitive, and rapid on-site detection of ASFV at the RNA level in the markets and farms.

Key words: aptamer, RNA, turn-on, African swine fever viru

Chengcheng Han, Kai Xia, Ruying Gong, Xuhan Wu, Lei Zhang, Xinle Liang. Design and application of lightening Spinach-p54 RNA apatmer to detect African swine fever virus[J]. Hereditas(Beijing), 2021, 43(12): 1170-1178.

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适配体名称	序列(5′→3′)
Spinach-p54-1	GATGCCGCGGATGGCGAAGGACGGGTCCAGTGCGTTCGCGCACTGTTGAGTAGAGTGTGAGCGCCAAACCGGTTGA
Spinach-p54-2	ACTGTCGTGTATGGCGAAGGACGGGTCCAGTGCGTTCGCGCACTGTTGAGTAGAGTGTGAGCGCCAGGCTCAGTCG
Spinach-p54-3	CCATAATGCCGTGGCGAAGGACGGGTCCAGTGCGTTCGCGCACTGTTGAGTAGAGTGTGAGCGCCCGGATAAACCG

RNA适配体名称	单链适配体最小自由能(kcal/mol)	与p54 RNA结合后最小自由能(kcal/mol)
Spinach-p54-1	-24.2	-52.88
Spinach-p54-2	-27.4	-51.38
Spinach-p54-3	-24.5	-46.28

Design and application of lightening Spinach-p54 RNA apatmer to detect African swine fever virus

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