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Journal of Tea Science ›› 2019, Vol. 39 ›› Issue (6): 669-680.doi: 10.13305/j.cnki.jts.2019.06.006

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Cloning and Expression Analysis of Serine Protease EoSP1 in Tea Geometrid (Ectropis obliqua) and Its Response to Starvation

ZHANG Xin1,2, Chen Chengcong3, DU Qin1,2, LI Xiwang1,2, SUN Xiaoling1,2,*   

  1. 1. Tea Research Institute of the Chinese Academy of Agricultural Sciences, Hangzhou 310008, China;
    2. Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture and Rural Affairs, Hangzhou 310008, China;
    3. National Research Center of Engineering and Technology of Tea Quality and Safety, Quanzhou 362400, China
  • Received:2019-10-08 Revised:2019-10-23 Online:2019-12-15 Published:2019-12-24

Abstract: Serine protease plays an important role in the digestion process of Lepidoptera insects. In this study, we cloned a serine protease encoding gene EoSP1 from Ectropis obliqua and analyzed its basic characteristics and expression patterns. The coding sequence of EoSP1 is 858 bp, encoding 285 amino acid residues with deduced molecular weight of 29.53 kDa and isoelectric point of 5.44. Compared with other serine proteases, EoSP1 contains conserved serine protease catalytic sites (H95, A161 and S328) and protein interaction domains, and shows the closest relationship with SPs from Mamestra configurata. Further, EoSP1-GST fusion protein similar to the predicted size was purified from E. coli cells. qRT-PCR analysis showed that the expression level of EoSP1 was much higher in larvae than that in adults, pupae and eggs, and expressed in midgut of larvae specifically. EoSP1 was down-regulated by starvation treatment, and the expression level was change back to that of control group after re-feeding. The above results provide a basis for the function analysis of digestive enzyme and screening of new insect-resistance targets in Ectropis obliqua.

Key words: Ectropis obliqua, serine protease, cloning, expression pattern

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