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  • 标题:Reference gene selection for quantitative real-time polymerase chain reaction analysis in Bombyx mori nucleopolyhedrovirus-infected silkworms
  • 本地全文:下载
  • 作者:Z Nie ; P Lü ; X Chen
  • 期刊名称:Invertebrate Survival Journal
  • 电子版ISSN:1824-307X
  • 出版年度:2017
  • 卷号:14
  • 期号:1
  • 页码:94-102
  • DOI:10.25431/1824-307X/isj.v14i1.94-102
  • 语种:English
  • 出版社:Universita degli Studi di Modena e Reggio Emilia
  • 摘要:Bombyx mori nucleopolyhedrovirus (BmNPV) is the most serious viral disease in silkworms. To investigate the mechanisms of the immune responses of B. mori to a BmNPV infection, a suitable reference gene (RG) is necessary for normalizing data when studying the expression of genes in BmNPV-infected silkworms or cells. Thus, quantitative real-time PCR polymerase chain reaction was used to compare the stability of expression of nine potential RGs, including the actin A3, translation initiation factor 3 (TIF-3), TIF-A4, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S RNA, 28S RNA, TATA-binding protein (TBP), ribosomal protein L3 (Rpl3), and α-tubulin genes, in silkworms infected with BmNPV. The results were analyzed by BestKeeper, geNorm, and NormFinder software. Overall, α-tubulin exhibited the most stable gene expression in BmNPV-infected silkworms, and this was verified by western blotting of the α-tubulin protein. Moreover, we detected the expression of some genes involved in the immune signaling pathways of silkworms after BmNPV infection using α-tubulin as an internal RG.
  • 关键词:Bombyx mori;reference genes;BmNPV;qPCR;immune genes
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