Molecular cloning and expression of the chemosensory Plutella xylostella protein gene, PxylCSP5
Author of the article:LI Fang-Yuan；FU Shu-Hui；ZHANG Bo-Chen；LV Zhi-Shen；WANG Hai-Qi；YAN Xi-Zhong；HAO Chi
Author's Workplace：College of Plant Protection, Shanxi Agricultural University, Taigu 030801, China；College of Plant Protection, Shanxi Agricultural University, Taigu 030801, China
Key Words：Plutella xylostella; chemosensory protein; gene cloning; spatio-temporal expression; prokaryotic expression
Abstract：
[Objectives] To investigate the function of the
chemosensory protein PxylCSP5 in Plutella xylostella. [Methods] The PxylCSP5 gene was cloned
using RT-PCR, and its sequence analyzed using bioinformatics methods. RT-qPCR was used to analyze the
expression of the PxylCSP5 gene in different developmental stages and
tissues of adult moths. A prokaryotic expression vector for PxylCSP5 was
constructed in BL21(DE3) [Results] The
complete open reading frame of PxylCSP5 is 393 bp, encodes 130 amino
acid residues, contains a signal peptide sequence of 18 amino acids at the
N-terminal and has four conserved cysteines. The
results of amino acid sequence alignment indicate that PxylCSP5 was
homologous to Helicoverpa zea HzeaCSP with 67.91% amino acid sequence
identify. RT-qPCR results show that PxylCSP5 was more highly expressed
in adults, and within adult tissues, in the legs. A prokaryotic expression
vector for PxylCSP5 was successfully constructed allowing PxylCSP5
to be expressed in vitro. [Conclusion] The PxylCSP5 gene is highly expressed
in the legs of adults, which suggests that its function may be related to host
location.