摘要
MDM2(murine double minute)是肿瘤抑制因子P53最重要的负性调控因子。为了解家蚕是否存在mdm2基因,利用人和小鼠的mdm2基因作为电子探针在家蚕EST数据库中检索到了同源序列。根据该序列设计引物,在家蚕蛹cDNA中扩增出开放阅读框长度为885 bp的mdm2同源基因,命名为Bm-mdm2(GenBank登录号:KF201646)。序列结构分析表明该基因编码的蛋白质序列具有MDM2保守域;同源性分析显示Bm-mdm2与赤拟谷盗(Tribolium castaneum)、大斑黑蝶(Danaus plexippus)mdm2的亲缘关系较近。为探讨Bm-mdm2与Bmp53的互作关系,利用家蚕核型多角体病毒(BmNPV)感染及H2O2刺激BmN细胞后,分析2个基因的表达变化。结果显示Bm-mdm2表达量下调,而Bmp53的表达量上调,并且Bmp53与Bm-mdm2的表达量变化存在相反的趋势,推测二者之间可能存在负调控关系。
Murine double minute （MDM2） is one of the most important negative regulators of tumor-suppressor protein P53. To investigate whether silkworm （ Bombyx mori） contains mdm2 gene, mdm2 gene sequences from Homo sapiens and Mus musculus were used as electron probes to search homologous genes in EST database of silkworm and a homol- ogous sequence was obtained. With the primers designed according to the obtained homologous sequence, an mdrn2- homologous gene with open reading frame of 885 bp was amplified from cDNA library constructed from silkworm pupa and named Bm-mdm2 （GenBank accession No. KF201646）. Sequence structure analysis revealed that protein se- quence encoded by this gene contained the conserved domains of MDM2. Homology analysis revealed that Bm-mdrn2 had a close genetic relationship with mdm2 from Tribolium castaneum and Culex quinquefasciatus. To explore the regula- tory relationship between Bm-mdrn2 and Bmp53, we analyzed the expressional variation of Bmp53 and Bmmdm2 in BmN cells after treatment with Bombyx moil nucleopolyhedrovirus （BmNPV） and H202. The results showed that the expres- sion level of Bm-mdrn2 was down-regulated while that of Bmp53 was up-regulated. Meanwhile, the expression levels of these two genes showed an opposite tendency, suggesting that a negati,Je regulatory relationship exists between Bmp53and Bm-mdm2.
出处
《蚕业科学》
CAS
CSCD
北大核心
2013年第5期887-892,共6页
ACTA SERICOLOGICA SINICA
基金
现代农业产业技术体系专项(No.CARS-22)
