Bao-2011-An immune-induced Re.pdf

lable at ScienceDirect Insect Biochemistry and Molecular Biology 41 (2011) 696e706Contents lists avaiInsect Biochemistry and Molecular Biology journal homepage: www.elsevier .com/locate/ ibmbAn immune-induced Reeler protein is involved in the Bombyx mori melanization cascade Yan-Yuan Bao, Jian Xue, Wen-Juan Wu, Ying Wang, Zu-Yao Lv, Chuan-Xi Zhang* Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Yuhangtang Road 866, Hangzhou, Zhejiang 310058, Chinaa r t i c l e i n f o Article history: Received 20 February 2011 Received in revised form 5 May 2011 Accepted 12 May 2011 Keywords: Silkworm Bombyx mori Immune-induced Reeler protein Nodulation response Phenoloxidase activityAbbreviations: PBS, phosphate-buffered saline; Prophenoloxidase; GFP, green fluorescent protein; Tn- 5B1-4; RACE, rapid amplification of cDNA ends; RN double-stranded RNA; Real-time qPCR, real-time quan transcription PCR; MCS, multiple cloning sites; OR glutathione-S-transferase; MWCO, molecular-weigh peroxidase; Ig, immunoglobulin; L-dopa, L-3,4-dihydr * Corresponding author. Tel./fax: t86 571 8898299 E-mail address: chxzhang@ (C.-X. Zhang 0965-1748/$ e see front matter
2011 Elsevier Ltd. doi:10.1016/j.ibmb.2011.05.001a b s t r a c t In this study, we isolated two reeler cDNAs from bacteria-challenged larval fat bodies of the silkworm, Bombyx mori. A reeler domain spanned most of the coding regions of these two cDNAs, and their expression patterns were different in B. mori larval tissues. The reeler1 gene was strongly induced by Escherichia coli K12 and Bacillus subtilis in B. mori larval hemocytes, fat bodies andmidguts, but reeler2was expressed at extremely low levels in these tissues. We focused on the reeler1 gene for functional analysis. Interference by double-stranded reeler1 RNA in vivo led to reduced nodule formation in bacteria-injected larvae, while the injection of recombinant Reeler1 promoted no