家蚕Tret1-X1基因对家蚕核型多角体病毒(BmNPV)的基因表达和基因组复制的影响  

Effects of Silkworm Tret1-X1 Gene on Gene Expression and Genome Replication of Bombyx mori Nucleopolyhedrovirus

作　　者：宋秋云 高娟 邵露璐 张业顺 方瑷[1,2] 张国政 Song Qiuyun;Gao Juan;Shao Lulu;Zhang Yeshun;Fang Ai;Zhang Guozheng(School of Biotechnology,Jiangsu University of Science and Technology,Zhenjiang Jiangsu 212100,China;Sericultural Research Institute,Chinese Academy of Agricultural Sciences,Zhenjiang Jiangsu 212100,China)

机构地区：[1]江苏科技大学生物技术学院,江苏镇江212100 [2]中国农业科学院蚕业研究所,江苏镇江212100

出　　处：《蚕业科学》2023年第3期244-253,共10页ACTA SERICOLOGICA SINICA

基　　金：财政部和农业农村部国家现代农业产业技术体系项目。

摘　　要：家蚕促海藻糖转运蛋白1异构体X1(Bombyx mori facilitated trehalose transporter Tret1-like isoform X1,BmTret1-X1)主要参与家蚕糖代谢过程。为探究其在家蚕抗BmNPV感染过程中的作用,克隆BmNPV抗性家蚕品系AN与易感品系C108的BmTret1-X1基因,发现基因编码区存在4个SNP导致的氨基酸改变,两者的预测蛋白质结构存在差异。利用pIZT/V5-His-mCherry表达载体在BmN细胞中过表达BmTret1-X1,BmNPV感染后24 h、48 h的BmNPV增殖受到抑制,BmNPV基因lef-3、vp39、p10和gp64的转录水平低于对照组(P<0.05),其中感染后24 h vp39基因的表达量与对照组相比差异达1 089倍,细胞中病毒基因组DNA复制也受到明显抑制。转染BmTret1-X1基因siRNA使BmN细胞中该基因表达量降低约一半,BmNPV的lef-3、vp39、p10和gp64转录水平升高,但BmNPV增殖和基因组DNA复制水平未见明显变化。综上所述,BmN细胞中BmTret1-X1基因的高表达对BmNPV的基因表达、DNA复制及病毒增殖具有抑制作用,该基因在家蚕对BmNPV的抗性形成过程中具有一定作用。Bombyx mori facilitated trehalose transporter Tret1-like isoform X1(BmTret1-X1)is mainly involved in the sugar metabolism of silkworm.In order to study its role in anti-BmNPV infection to silkworm,BmTret1-X1 gene of the resistant strain AN and the susceptible strain C108 were cloned in this study.It was found that there were amino acid changes caused by four SNPs in their gene coding region,and some differences in their predicted protein structures.With the overexpression of BmTret1-X1 in BmN cells using the pIZT/V5-His-mCherry expression vector,the proliferation of BmNPV at 24 h and 48 h post infection was inhibited,and the transcription levels of BmNPV genes lef-3,vp39,p10,and gp64 were lower than those in the control group(P<0.05).The expression of vp39 gene at 24 h post infection was 1089 times higher than that in the control group,and the viral genomic DNA replication in BmN cells was also significantly inhibited.Transfection of the BmTret1-X1 gene siRNA reduced the expression level of this gene in BmN cells by about half,increased the transcription levels of BmNPV lef-3,vp39,p10,and gp64,but there was no significant effect on BmNPV proliferation and genomic DNA replication levels.In summary,the high expression of BmTret1-X1 gene in BmN cells has an inhibitory effect on the gene expression,DNA replication and viral proliferation of BmNPV,and the gene plays a certain role in the formation of BmNPV resistance in silkworm.

关 键 词：家蚕 BmTret1-X1基因 BmNPV抗性 病毒复制 病毒基因表达 

分 类 号：S884.51[农业科学—特种经济动物饲养]