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Your Position :Home->Past Journals Catalog->2015年52 No.3

Cloning and expression analysis of the polycalin gene in the cotton bollworm Helicoverpa armigera
Author of the article:WANG Ya-Nan1, 2** ZHONG Feng1 WEI Ji-Zhen1 XIE Bing-Tang1ZHANG Wan-Na1 CHEN Li-Zhen2***LIANG Ge-
Author's Workplace:1. College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China; 2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Chinese Academy of Agriculture Science, Beijing 100193, China
Key Words:Helicoverpa armigera, polycalin gene, expressin, Bt toxin
Abstract: [Objetives]  To further define the role of Polycalin in the Cry1Ac resistance mechanism of Helicoverpa armigera in and thereby develop a feasible resistance management strategy for this pest. [Methods]  The full-lengthsequence of the polycalin coding gene was obtained using RT-PCR (reverse transcription-polymerase chain reaction) and RACE (rapid amplification of cDNA ends) technology. The expression levels of polycalin in different development stages and different tissues of the larval digestive tract were analyzed using real-time quantitative PCR. Changes in the expression of polycalin in the larval midgut in 4th instar larvae fed on an artificial diet containing the Bt insecticide protein Cry1Ac were also compared. [Results]  The results indicate that the full-length of the polycalin from H. armigera coding gene was 2 955 bp (GenBank accession number KP100652), the open reading frame was 2 781 bp in length, encoding 926 amino acid residues. The predicted molecular weight was 101.68 ku and the isoelectric point was 4.95. The putative protein sequence contained a N-terminal signal peptide of 20 amino acids, three N-linked and eight O-linked glycosylation sites, and a GPI anchor signal peptide with 2 amino acids at the C-terminus. Polycalin was expressed in all growth stages of H. armigera, and expression in larvae was higher than that in other stages, especially in 1st-3rd instar larvae. The lowest expression levels occurred in eggs, adults and pupae. The expression of the polycalin gene was dramatically suppressed after larvae were fed on food containing the Cry1Ac toxin. [Conclusion]  These results provide a basis for further clarifying the function of polycalin with respect to the Bt toxin.

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