Recovery and genetic characterization of black queen cell virus

被引:2
|
作者
Yang, Sa [1 ,2 ,3 ]
Deng, Yanchun [4 ]
Zhang, Li [1 ,2 ]
Wang, Xinling [1 ,2 ]
Deng, Shuai [4 ]
Dai, Pingli [1 ,2 ]
Hou, Chunsheng [4 ]
机构
[1] Chinese Acad Agr Sci, Inst Apicultural Res, Beijing, Peoples R China
[2] Minist Agr, Key Lab Pollinating Insect Biol, Beijing, Peoples R China
[3] Chinese Acad Agr Sci, Grad Sch, Beijing 100081, Peoples R China
[4] Chinese Acad Agr Sci, Inst Bast Fiber Crops, Changsha 410205, Peoples R China
来源
JOURNAL OF GENERAL VIROLOGY | 2022年 / 103卷 / 08期
基金
中国国家自然科学基金;
关键词
Apis mellifera; black queen cell virus; phylogenetic analysis; infectious clone; reverse genetic system; expression vector; COMPLETE GENOME SEQUENCE; CHINESE SACBROOD VIRUS; APIS-MELLIFERA; INFECTED HONEYBEES; BEE VIRUSES; IDENTIFICATION; REGION; CERANA;
D O I
10.1099/jgv.0.001770
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Black queen cell virus (BQCV) is a severe threat to the honeybee (Apis mellifera) worldwide. Although several BQCV strains have been reported in China, the molecular basis for BQCV pathogenicity has not been well understood. Thus, a reverse genetic system of BQCV is required for studying viral replication and its pathogenic mechanism. Here, the complete genome sequence of BQCV was obtained from honeybees using reverse transcription PCR (RT- PCR), namely a BQCV China- GS1 strain (KY741959). Then, a phylogenetic tree was built to analyse the genetic relationships among BQCV strains from different regions. Our results showed that the BQCV China- GS1 contained two ORFs, consistent with the known reference strains, except for the BQCV China-JL1 strain (KP119603). Furthermore, the infectious clone of BQCV was constructed based on BQCV China- GS1 using a low copy vector pACYC177 and gene recombination. Due to the lack of culture cells for bee viruses, we infected the healthy bees with infectious clone of BQCV, and the rescued BQCV resulted in the recovery of recombinant virus, which induced higher mortality than those of the control group. Immune response after inoculated with BQCV further confirmed that the infectious clone of BQCV caused the cellular and humoral immune response of honeybee (A. mellifera). In conclusion, the full nucleotide sequence of BQCV China- GS1 strain was determined, and the infectious clone of BQCV was constructed in this study. These data will improve the understanding of pathogenesis and the host immune responses to viral infection.
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页数:11
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