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Expression of anti-NbHK single-chain antibody in fusion with NSlmb enhances the resistance to Nosema bombycis in Sf9-III cells

Published online by Cambridge University Press:  06 April 2022

Renze Zhang
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China
Shiyi Zheng
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Affiliated Jinhua Hospital, Zhejiang University of Medicine, Jinhua Municipal Central Hospital, Jinhua, Zhejiang 321000, China
Hongyun Huang
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China
Xi Sun
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China
Yukang Huang
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China
Junhong Wei
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University, Chongqing 400715, China
Guoqing Pan
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University, Chongqing 400715, China
Chunfeng Li*
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University, Chongqing 400715, China
Zeyang Zhou
Affiliation:
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China Chongqing Key Laboratory of Microsporidia Infection and Control, Southwest University, Chongqing 400715, China College of Life Sciences, Chongqing Normal University, Chongqing 401331, China
*
Author for correspondence: Chunfeng Li, Email: [email protected]

Abstract

Nosema bombycis is a destructive and specific intracellular parasite of silkworm, which is extremely harmful to the silkworm industry. N. bombycis is considered as a quarantine pathogen of sericulture because of its long incubation period and horizontal and vertical transmission. Herein, two single-chain antibodies targeting N. bombycis hexokinase (NbHK) were cloned and expressed in fusion with the N-terminal of Slmb (a Drosophila melanogaster FBP), which contains the F-box domain. Western blotting demonstrated that Sf9-III cells expressed NSlmb–scFv-7A and NSlmb–scFv-6H, which recognized native NbHK. Subsequently, the NbHK was degraded by host ubiquitination system. When challenged with N. bombycis, the transfected Sf9-III cells exhibited better resistance relative to the controls, demonstrating that NbHK is a prospective target for parasite controls and this approach represents a potential solution for constructing N. bombycis-resistant Bombyx mori.

Type
Research Paper
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press

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Footnotes

*

These authors contributed equally to the work reported in this paper.

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