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Cloning and characterization of two glutathione S ‐transferase cDNAs in the spruce budworm, Choristoneura fumiferana
Author(s) -
Zheng Sichun,
Deng Huimin,
Ladd Tim,
Tomkins Bill L.,
Krell Peter J.,
Feng Qili
Publication year - 2007
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.20206
Subject(s) - biology , midgut , choristoneura fumiferana , spruce budworm , complementary dna , microbiology and biotechnology , glutathione s transferase , cdna library , gene expression , gene , moulting , peptide sequence , hevea brasiliensis , biochemistry , glutathione , botany , tortricidae , enzyme , larva , chemistry , natural rubber , organic chemistry
Two Choristoneura fumiferana glutathione S ‐transferase cDNAs were cloned from a cDNA library constructed using mRNA from the midgut cell line, CF‐203. These cDNAs ( Cf GST2, Cf GST3) encoded two structurally different proteins with a predicted molecular mass of 21 and 24 kDa, respectively, which was confirmed through protein expression in a bacterial system. Quantitative reverse‐transcription PCR analyses revealed that the transcripts of these two genes were present in the epidermis, fat body, and midgut of the 6th instar larvae. Cf GST2 was expressed in the fat body when the insects were close to pupal molting, while it was constantly expressed in the other two tissues during the 6th instar stage. Cf GST3 gene was expressed highly and constantly in all of the tissues throughout the 6th instar stage. Immunohistochemistry analysis demonstrated that Cf GST2 and Cf GST3 proteins were present mainly in the fat body and epidermis and no protein was detected in the midgut. Cf GST2 and Cf GST3 were different from Cf GST reported before (Feng et al., 1999: Insect Biochem Mol Biol 29:779–793) in amino acid sequence, expression pattern, and responsiveness to tebufenozide. Arch. Insect Biochem. Physiol. 66:146–157, 2007. © 2007 Wiley‐Liss, Inc.