Cloning and characterization of two glutathione S-transferase cDNAs in the spruce budworm, Choristoneura fumiferana.
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 (CfGST2, CfGST3) 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. CfGST2 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. CfGST3 gene was expressed highly and constantly in all of the tissues throughout the 6th instar stage. Immunohistochemistry analysis demonstrated that CfGST2 and CfGST3 proteins were present mainly in the fat body and epidermis and no protein was detected in the midgut. CfGST2 and CfGST3 were different from CfGST reported before (Feng et al., 1999: Insect Biochem Mol Biol 29:779-793) in amino acid sequence, expression pattern, and responsiveness to tebufenozide.
Zheng
Sichun
S
Guangdong Provincial Key Lab of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou, China.
Deng
Huimin
H
Ladd
Tim
T
Tomkins
Bill L
BL
Krell
Peter J
PJ
Feng
Qili
Q
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D005982:Glutathione Transferase / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D006834:Hydrazines / Q000494:pharmacology; D019476:Insect Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009036:Moths / Q000201:enzymology* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein
D018076:DNA, Complementary; D006834:Hydrazines; D019476:Insect Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; C082026:tebufenozide