Full length cDNA cloning and expression of the chitin synthasegene in Tetranychus cinnabarinus
Author of the article:XIN Tian-Rong** ZOU Zhi-Yong MIN Qiang LI Lei WU Xuan-DeWANG Ya-Yu ZOU Zhi-Wen XIA Bin***
Author's Workplace：College of Life Science, Nanchang University, Nanchang 330031, China
Key Words：Tetranychus cinnabarinus, chitin synthase, gene cloning, expression
Abstract： [Objectives] To clone the Tetranychus cinnabarinus chitin synthase gene and detect relative levels of the expression of this gene in the different developmental stages of T. cinnabarinus. [Methods] The full-length chitin synthase cDNA of T. cinnabarinus was first cloned by RT-PCR and RACE-PCR, and the relative expression levels of the TcCHS1 gene were then detected in different developmental stages of T. cinnabarinus using quantative real-time PCR. [Results] TcCHS1 cDNA was 4 881 bp in length, including a 198 bp 5¢ terminal UTR, 4 425 bp encoding region comprised of 1 474 amino acids and a 258 bp 3¢ terminal UTR. The theoretical molecular weight of TcCHS1 based on the deduced amino acid sequence was estimated to be 168.35 ku, with an isoelectric point of 6.26. The signature sequences “EDR” and “QRRRW” for chitin synthases were found in TcCHS1. Alignment of the deduced amino acid sequence of TcCHS1 and those of other insects demonstrated that this shared 98% and 55% similarity with T. urticae and Metaseiulus occidentalis, respectively, and approximately 50% similarity with other insects. Phylogenetic tree analysis revealed that the putative CHS amino acid sequence of T. cinnabarinus showed high similarity with the CHS1 sequences of other insects, and was most closely related to that of T. urticae. Real-time quantitative PCR demonstrated that the TcCHS1 gene was expressed in different development stages, including eggs, larvae, protonymphs, deutonymphs, adult females and adult males. The result suggest that the TcCHS1 mRNA were most highly expressed in eggs and adult females, and least expressed in deutonymphs. [Conclusion] The results indicate that TcCHS1 may play an important role in the growth and development of T. cinnabarinus.