Cloning, characterization and expression of two glutathione S-transferase cDNAs in the spruce budworm, Choristoneura fumiferana.
Two Choristoneura fumiferana glutathione S-transferase cDNAs (CfGSTs4 and CfGSTd5) were cloned from a cDNA library constructed using mRNA from the midgut cell line, CF-203. These cDNAs encoded two structurally different proteins with a predicted molecular mass of 23 and 24 kDa, respectively. Amino acid sequence analysis indicates that CfGSTs4 and CfGSTd5 contained Sigma and Delta GST domain, respectively. CfGSTs4 cDNA was expressed as a recombinant protein with the same molecular mass as predicted. Semi-quantitative reverse-transcription PCR analyses indicated that both of these genes were expressed in the epidermis, fat body, and midgut of the 6th instar larvae, as well as CF-203 cells. CfGSTs4 was highly and almost constantly expressed in all tissues during the 6th instar stage. There were higher levels of CfGSTs4 protein in the midgut and fat body than in the epidermis. CfGSTd5 was expressed in the fat body when the insects underwent pupal molting and was constantly expressed in the epidermis and midgut during 6th instar development. CfGSTs4 expression was not affected by ecdysone agonist tebufenozide (RH5992), whereas CfGSTd5 expression was slightly suppressed by the compound.
Huang
Yufen
Y
Guangdong Provincial Key Lab of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou, China.
Krell
Peter J
PJ
Ladd
Tim
T
Feng
Qili
Q
Zheng
Sichun
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D007527:Isoenzymes; D007814:Larva / Q000254:growth & development / Q000378:metabolism; D008969:Molecular Sequence Data; D009036:Moths / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny; D011679:Pupa / Q000254:growth & development / Q000378:metabolism
D018076:DNA, Complementary; D007527:Isoenzymes; D005982:Glutathione Transferase