Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (8): 113-119.

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RNA Interference of Antifreeze Protein Gene in Tenebrio molitor Mediated by Bacterially Expressed dsRNA

Shi Meng, Liu Xiaoning, Ma Ji   

  1. Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046
  • Revised:2014-01-18 Online:2014-08-15 Published:2014-08-01
  • Contact: 通讯作者: 马纪,博士,教授,研究方向:昆虫低温分子生物学;E-mail:majiuci@xju.edu.cn
  • Supported by:
    国家自然科学基金项目(31360527);新疆生物资源基因工程重点实验室开放课题(XJDX0201-2014-03)

Abstract: In order to further study the non-freezing function of antifreeze protein(AFP)genes in Tenebrio molitor using RNAi technology, the RNAi fragment of Tmafp433 from T. molitor was inserted into L4440 dsRNA expression vector, and transformed into E. coli HT115(DE3). The dsRNA corresponding to Tmafps, designated as Tmafp-dsRNA, was expressed by IPTG induction, and injected into the larvae after purification. The mRNA level of Tmafps was detected using real-time quantitative PCR. The results showed that the E. coli HT115(DE3)containing L4440-Tmafp recombinant plasmid can express Tmafp-dsRNA. After 24 h of injection, the expression of Tmafps in T. molitor was significantly decreased to 60.8% of the control, suggesting that the expression of Tmafps was inhibited by injecting the bacterially expressed Tmafp-dsRNA. The present results laid foundation for further research in afp gene function.

Key words: Tenebrio molitor, afp gene, RNA interference, Double-stranded RNA, L4440 vector