家蚕核型多角体病毒极早期蛋白PE38与家蚕SRSF蛋白激酶的相互作用关系验证

Verification of Interaction Between BmNPV Immediate-early Protein PE38 and BmSRPK of Silkworm,Bombyx mori

家蚕核型多角体病毒(Bm NPV)是家蚕血液型脓病的病原,其极早期蛋白PE38参与病毒的侵染、复制与增殖。通过酵母双杂交和荧光双分子互补(Bi FC)试验证实Bm NPV PE38蛋白和家蚕SRSF蛋白激酶(Bombyx mori SRSF protein kinase 1-like,Bm SRPK)之间存在互作关系,推测Bm NPV可能是通过影响宿主的前体mRNA组成性和选择性剪接来获得对自身增殖有利的微环境。进一步利用实时荧光定量PCR(qRT-PCR)检测分析感染Bm NPV后Bm N细胞中Bm SRPK的表达变化以及正常家蚕5龄第3天幼虫体内Bm SRPK表达的组织特异性,发现感染病毒后Bm N细胞中Bm SRPK的表达量有所增加;Bm SRPK在正常幼虫的精巢、中肠和脂肪体中表达量较高,在血细胞中的表达量最低。研究结果有助于进一步深入研究Bm NPV和家蚕宿主之间复杂的相互作用关系。

Bombyx mori nuclopolyhedrovirus（Bm NPV）is the causative agent of silkworm（Bombyx mori）nuclopolyhedrosis disease.Its immediate-early protein PE38 has a significant impact on infectivity,replication and reproduction of the virus.In this study,we confirmed that Bm NPV PE38 can interact with Bombyx mori SRSF protein kinase（BmSRPK）in silkworm using yeast two-hybrid system and bimolecular fluorescence complementation system.It is suggested that Bm NPV creates favorable micro-environment through influencing constitutive and selective splicing of host pre-mRNAs.Furthermore,expressional change of Bm SRPK gene in Bm N cells after Bm NPV infection and tissue-specific expression profiles of Bm SRPK in day 3 silkworm larvae of the 5th instar were analyzed by quantitative real-time PCR（qRT-PCR）.It was found that expression of Bm SRPK gene in Bm N cells was increased after Bm NPV infection,that in larval testis,midgut and fat body was relatively higher,and that in hemocyte was the lowest.These results are conducive to further investigations on the complicated interaction between Bm NPV and its host silkworm.