›› 2015, Vol. 58 ›› Issue (2): 115-121.

• RESEARCH PAPERS • Previous Articles     Next Articles

Gene expression profiling and polyclonal antibody preparation of the transcription factor c-Myc in the brain of diapause and non-diapause pupae of the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuiadae)

CHEN Wei1,*, XU Wei-Hua2   

  1. (1. Guangdong Province Key Laboratory for Biotechnology Drug Candidates, School of Biosciences and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou 510006, China; 2. State Key Laboratory of Biocontrol and Institute of Entomology, School of Life Sciences, Sun YatSen University, Guangzhou 510275, China)
  • Online:2015-02-20 Published:2015-02-20

Abstract: 【Aim】 c-Myc is an intensively studied transcription factor and also an important downstream target of Wnt/β-catenin signaling pathway. The objective of this research was to clone the c-myc cDNA from Helicoverpa armigera , to investigate the expression of Har-c-myc in mRNA level, and to prepare polyclonal antibody against Har-c-Myc. 【Methods】 Har-c-myc was cloned by RACE. The mRNA levels of Har-c-myc in the brain of non-diapause and diapause pupae were investigated by RT-PCR. Prokaryotic expression vector of  Har-c-myc was constructed and the recombinant protein was expressed in Escherichia coli . The purified recombinant protein was injected into a New Zealand rabbit to generate polyclonal antibody. The antibody titer was determined by ELISA. 【Results】 Har-c-myc cDNA was cloned successfully from H. armigera. The mRNA levels of Har-c-Myc were significantly lower in the brain of diapause pupae than in the brain of non-diapause pupae. Recombinant Har-c-Myc was successfully expressed in E. coli and purified by Ni-NTA agarose column. The titer of the antibody against Har-c-Myc was estimated by ELISA as high as 1:125 000. 【Conclusion】 The expression of Har-c-myc is down-regulated in the brain of diapause pupae. Antibody against Har-c-Myc was obtained. This study lays a foundation for further investigating the function of Wnt/β-catenin in the diapause of H. armigera.

Key words: Helicoverpa aimigera, diapause, transcription factor, c-Myc, polyclonal antibody, Wnt/β-catenin signaling pathway