Cloning and Expression Analysis of the Positive Regulatory Domain Zinc Finger Protein 10 Like Gene of Helicoverpa armigera
ZHANG Ya-Lin, HU De-Qin, LI Yuan, HUANGFU Hui-Lin, JIAENLISINI•Na-Zha-Er, YU Rui, LIU Xiao-Ning*
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering/College of Life Science and Technology, Xinjiang University, Urumqi 830046, China
Abstract：Helicoverpa armigera is an omnivorous pest that causes severe yield loss of many crops. The current planting of Bt crops and the extensive use of chemical pesticides have enabled the cotton bollworm to evolve a defense mechanism that protects itself against pesticides and plants secondary substances. In order to develop an environment friendly insecticide for harmless control of H. armigera, the core sequence HE1, which can respond to 2-tridecanone (2-TD), of the cytochrome P450 CYP6B6 promoter was used as a probe to screen out a positive regulatory domain zinc finger protein 10 like (PRDM10l) gene by using DNA pull down technology. The sequence of HaPRDM10l were cloned from 6th instar larvae of H. armigera, analysed by bioinformatics and its fusion protein expressed by using a prokaryotic expression system. qPCR was used to detect the spatial-temporal expression profile of HaPRDM10l in H. armigera, and its expression pattern in the midgut of the 6th instar larvae of H. armigera was detected after 2-TD treatment. The results showed that the ORF of HaPRDM10l (GenBank No XM_021334552.1) was 2 067 bp, encoded 688 amino acids. The molecular weight and theoretical isoelectric point of the protein were 77.96 kD and 8.84, respectively. HaPRDM10l was expressed at all developmental stages of H. armigera, with the highest expression in 5th instar larvae, and among different tissues the highest expression in the midgut of 6th instar larvae. After the cotton bollworm was treated with different concentrations of 2-TD, both HaPRDM10l and CYP6B6 could respond to the induction of 2-TD. The expression of HaPRDM10l reached the maximum when 10 mg/g of 2-TD treatment for 12 h, which was 7.6 folds of that in the control group, and the expression level of CYP6B6 reached the maximum at 6 h after 5 mg/g 2-TD treatment, which was 4.6 folds that of the control group. Further analysis revealed that there was a positive correlation between the expression of HaPRDM10l and CYP6B6 in the 6 h treatment group (r=0.749, P<0.01). The above results suggested that HaPRDM10l may regulate the expression of CYP6B6 in response to the stress of 2-TD. This study provides a reference for clarifying the mechanism of the H. armigera in response to the stress of plant secondary substances.
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