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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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Restriction maps of PxGSTe and PxGSTs genes from Plutella xylostella. Probes for PxGSTe and PxGSTs genes are shown under restriction maps as solid bars.
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f04: Restriction maps of PxGSTe and PxGSTs genes from Plutella xylostella. Probes for PxGSTe and PxGSTs genes are shown under restriction maps as solid bars.

Mentions: Total RNA (20 µg) isolated from whole body extracts of different developmental stages were size-fractionated on a 1.2% agarose gel containing 0.66 M formaldehyde and transferred to a Biodyne PLUS membrane (Pall Corp., www.pall.com). The blot was hybridized with a random-primed 32P-labeled fragment containing the coding sequences of PxGSTe or PxGSTs gene (Figure 4) (Sambrook et al. 1989).


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)

Restriction maps of PxGSTe and PxGSTs genes from Plutella xylostella. Probes for PxGSTe and PxGSTs genes are shown under restriction maps as solid bars.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f04: Restriction maps of PxGSTe and PxGSTs genes from Plutella xylostella. Probes for PxGSTe and PxGSTs genes are shown under restriction maps as solid bars.
Mentions: Total RNA (20 µg) isolated from whole body extracts of different developmental stages were size-fractionated on a 1.2% agarose gel containing 0.66 M formaldehyde and transferred to a Biodyne PLUS membrane (Pall Corp., www.pall.com). The blot was hybridized with a random-primed 32P-labeled fragment containing the coding sequences of PxGSTe or PxGSTs gene (Figure 4) (Sambrook et al. 1989).

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