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Cloning and
expression pattern of chitin synthase (CHS) gene in
epidermis of Ectropis obliqua Prout
Yue-Rong Liang1, Chen Lin1,
Ruo-Ren Wang1, Jian-Hui Ye1,
Jian-Liang Lu2*
1Zhejiang
University Tea Research Institute, Hangzhou 310029, PR
China.
2The
State Agriculture Ministry Laboratory of Horticulture Plant
Growth Development and Quality Improvement, Hangzhou 310029,
PR China.
*Corresponding author. E-mail:
jllu@zju.edu.cn.
Fax: 86 571 8697 1704.
Abbreviation: CHS,
Chitin
synthase; PTGS, post-transcriptional gene silencing;
RNAi, RNA interference; RACE, rapid
amplification of cDNA ends; TMH, transmembrane
helices; RT-PCR, reverse transcription polymerase
chain reaction; NPV, nucleopolyhedrovirus.
Accepted 25 June, 2010 |
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Ectropis obliqua
Prout is a major pest in tea fields. Chitin synthase (CHS)
plays an important role in biosynthesis of chitin and growth
of the pest. A cDNA sequence encoding the CHS and its
expression pattern during development of
E.obliqua
was investigated. The CHS cDNA sequence was 5496 bp
nucleotides, with an open reading frame of 4692 bp encoding
a protein of 1563 amino acids. It belonged to CHS-A member
of CHS gene family. Alternative splice was found in the
CHS-A cDNA and the alternatively spliced fragments were 177
bp and shared 65% identity with each other at the nucleotide
level. The CHS-A expression was the strongest in the third
and fourth instar larvae, during which the growth rate of
E. obliqua larvae was the rapidest. Catalysis model of
CHS-A enzyme in E. obliqua was also hypothesized
according to the specific motifs and topological structure
prediction of the protein. This study provided an important
information for further research on development of
RNA interference (RNAi) technology to control E. obliqua.
Key words:
Ectropis oblique,
tea pest, chitin synthase, gene cloning, RNAi, biological
control. |
