Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (9): 1808-1817.doi: 10.3864/j.issn.0578-1752.2013.09.008

• PLANT PROTECTION • Previous Articles     Next Articles

Molecular Cloning, Sequence Analysis and Expression Pattern Detection of α-Tubulin Gene from Helicoverpa armigera (Hübner)

 YAN  Shuo, ZHU  Jia-Lin, ZHU  Wei-Long, PAN  Li-Long, ZHANG  Qing-Wen, LIU  Xiao-Xia   

  1. Department of Entomology, College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193
  • Received:2013-01-04 Online:2013-05-01 Published:2013-02-17

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Abstract: 【Objective】The objective of this study is to clone and analyze a novel cDNA, named as HeTubA, encoding the α-tubulin from Helicoverpa armigera (Hübner), and detect the relative expression levels of HeTubA and HeTubB by real time PCR (QRT-PCR). 【Method】The total RNA was extracted from H. armigera 3rd instar larvae, and α-tubulin gene was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The relative expression levels of HeTubA and HeTubB were detected in different adult organs and developmental stages by QRT-PCR.【Result】Sequencing and structural analysis showed that the ORF of HeTubA was 1 353 bp in size, encoding 450 amino acid residues (GenBank accession number JQ069957). HeTubA contained conserved residues of α-tubulin. The homologue analysis revealed that HeTubA shared high identity with Xestia c-nigrum, Papilio xuthus and Bombyx mori α-tubulin, which indicated that insect α-tubulin was conserved in evolutional process. QRT-PCR revealed that HeTubA and HeTubB mRNA expression was neither adult organ-specific nor developmental-stage-specific. Relative expression levels of HeTubA and HeTubB were the highest in compound eye, the lowest in abdomen. Expression of HeTubA peaked at last larval instar and pupal development, whereas that of HeTubB peaked at last larval instar and adult development.【Conclusion】HeTubA was cloned and mRNA expression levels of HeTubA and HeTubB were detected. 3D structural of HeTubA was constructed. The study is helpful for further researches on the function of HeTubA and HeTubB and development of a new-type pesticide.

Key words: Helicoverpa armigera , α-tubulin , β-tubulin , gene cloning , gene expression , 3D structure

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