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Genomic organization and developmental expression of glutathione S-transferase genes of the diamondback moth, Plutella xylostella.

Sonoda S, Ashfaq M, Tsumuki H - J. Insect Sci. (2006)

Bottom Line: Based on amino acid identity with GST genes from other insects, this gene was classified as a member of the Sigma class.Southern blot analysis showed that PxGSTs was a single copy gene, whereas there were homologous members of the PxGSTe gene in the P. xylostella genome.RNA gel blot analysis indicated that the expression levels of both genes changed with the developmental stage of P. xylostella.

View Article: PubMed Central - PubMed

Affiliation: Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan. sonodas@rib.okayama-u.ac.jp

ABSTRACT
In the present study, we cloned and sequenced the entire coding regions of two glutathione S-transferase (GST) genes encoding PxGSTs and PxGSTe from the diamondback moth, Plutella xylostella L. (Lepidoptera: Yponomeutidae), along with their respective 5' and 3' flanking regions. The PxGSTs gene was composed of four exons and three introns. Based on amino acid identity with GST genes from other insects, this gene was classified as a member of the Sigma class. The gene encoding PxGSTe had an intron in the 5' flanking region. Southern blot analysis showed that PxGSTs was a single copy gene, whereas there were homologous members of the PxGSTe gene in the P. xylostella genome. RNA gel blot analysis indicated that the expression levels of both genes changed with the developmental stage of P. xylostella.

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Developmental expression of PxGSTe and PxGSTs genes from Plutella xylostella. Total RNA extracted from 1st and 2nd instar larvae (lane 1), 3rd instar larvae (lane 2), male 4th instar larvae (lane 3), female 4th instar larvae (lane 4), male pupae (lane 5), female pupae (lane 6), male adults (lane 7) and female adults (lane 8) were analyzed by RNA gel blot analysis. The photographs of the ethidium bromide-stained RNA gel before transfer are also shown.
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f06: Developmental expression of PxGSTe and PxGSTs genes from Plutella xylostella. Total RNA extracted from 1st and 2nd instar larvae (lane 1), 3rd instar larvae (lane 2), male 4th instar larvae (lane 3), female 4th instar larvae (lane 4), male pupae (lane 5), female pupae (lane 6), male adults (lane 7) and female adults (lane 8) were analyzed by RNA gel blot analysis. The photographs of the ethidium bromide-stained RNA gel before transfer are also shown.

Mentions: Developmental expression of PxGSTe gene was examined by RNA gel blot analysis (Figure 6). First and second instar larvae accumulated mRNA at very high levels. The mRNA levels apparently decreased in third instar larvae, and increased in the fourth instar, but little mRNA was detected in pupae. In adults, the PxGSTs mRNA levels increased again.


Genomic organization and developmental expression of glutathione S-transferase genes of the diamondback moth, Plutella xylostella.

Sonoda S, Ashfaq M, Tsumuki H - J. Insect Sci. (2006)

Developmental expression of PxGSTe and PxGSTs genes from Plutella xylostella. Total RNA extracted from 1st and 2nd instar larvae (lane 1), 3rd instar larvae (lane 2), male 4th instar larvae (lane 3), female 4th instar larvae (lane 4), male pupae (lane 5), female pupae (lane 6), male adults (lane 7) and female adults (lane 8) were analyzed by RNA gel blot analysis. The photographs of the ethidium bromide-stained RNA gel before transfer are also shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2990328&req=5

f06: Developmental expression of PxGSTe and PxGSTs genes from Plutella xylostella. Total RNA extracted from 1st and 2nd instar larvae (lane 1), 3rd instar larvae (lane 2), male 4th instar larvae (lane 3), female 4th instar larvae (lane 4), male pupae (lane 5), female pupae (lane 6), male adults (lane 7) and female adults (lane 8) were analyzed by RNA gel blot analysis. The photographs of the ethidium bromide-stained RNA gel before transfer are also shown.
Mentions: Developmental expression of PxGSTe gene was examined by RNA gel blot analysis (Figure 6). First and second instar larvae accumulated mRNA at very high levels. The mRNA levels apparently decreased in third instar larvae, and increased in the fourth instar, but little mRNA was detected in pupae. In adults, the PxGSTs mRNA levels increased again.

Bottom Line: Based on amino acid identity with GST genes from other insects, this gene was classified as a member of the Sigma class.Southern blot analysis showed that PxGSTs was a single copy gene, whereas there were homologous members of the PxGSTe gene in the P. xylostella genome.RNA gel blot analysis indicated that the expression levels of both genes changed with the developmental stage of P. xylostella.

View Article: PubMed Central - PubMed

Affiliation: Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan. sonodas@rib.okayama-u.ac.jp

ABSTRACT
In the present study, we cloned and sequenced the entire coding regions of two glutathione S-transferase (GST) genes encoding PxGSTs and PxGSTe from the diamondback moth, Plutella xylostella L. (Lepidoptera: Yponomeutidae), along with their respective 5' and 3' flanking regions. The PxGSTs gene was composed of four exons and three introns. Based on amino acid identity with GST genes from other insects, this gene was classified as a member of the Sigma class. The gene encoding PxGSTe had an intron in the 5' flanking region. Southern blot analysis showed that PxGSTs was a single copy gene, whereas there were homologous members of the PxGSTe gene in the P. xylostella genome. RNA gel blot analysis indicated that the expression levels of both genes changed with the developmental stage of P. xylostella.

Show MeSH