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Characterization of cDNAs encoding serine proteases and their transcriptional responses to Cry1Ab protoxin in the gut of Ostrinia nubilalis larvae.
PLoS One. 2012; 7(8):e44090.Plos

Abstract

Serine proteases, such as trypsin and chymotrypsin, are the primary digestive enzymes in lepidopteran larvae, and are also involved in Bacillus thuringiensis (Bt) protoxin activation and protoxin/toxin degradation. We isolated and sequenced 34 cDNAs putatively encoding trypsins, chymotrypsins and their homologs from the European corn borer (Ostrinia nubilalis) larval gut. Our analyses of the cDNA-deduced amino acid sequences indicated that 12 were putative trypsins, 12 were putative chymotrypsins, and the remaining 10 were trypsin and chymotrypsin homologs that lack one or more conserved residues of typical trypsins and chymotrypsins. Reverse transcription PCR analysis indicated that all genes were highly expressed in gut tissues, but one group of phylogenetically-related trypsin genes, OnTry-G2, was highly expressed in larval foregut and midgut, whereas another group, OnTry-G3, was highly expressed in the midgut and hindgut. Real-time quantitative PCR analysis indicated that several trypsin genes (OnTry5 and OnTry6) were significantly up-regulated in the gut of third-instar larvae after feeding on Cry1Ab protoxin from 2 to 24 h, whereas one trypsin (OnTry2) was down-regulated at all time points. Four chymotrypsin and chymotrypsin homolog genes (OnCTP2, OnCTP5, OnCTP12 and OnCTP13) were up-regulated at least 2-fold in the gut of the larvae after feeding on Cry1Ab protoxin for 24 h. Our data represent the first in-depth study of gut transcripts encoding expanded families of protease genes in O. nubilalis larvae and demonstrate differential expression of protease genes that may be related to Cry1Ab intoxication and/or resistance.

Authors+Show Affiliations

Department of Entomology, Kansas State University, Manhattan, Kansas, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22952884

Citation

Yao, Jianxiu, et al. "Characterization of cDNAs Encoding Serine Proteases and Their Transcriptional Responses to Cry1Ab Protoxin in the Gut of Ostrinia Nubilalis Larvae." PloS One, vol. 7, no. 8, 2012, pp. e44090.
Yao J, Buschman LL, Oppert B, et al. Characterization of cDNAs encoding serine proteases and their transcriptional responses to Cry1Ab protoxin in the gut of Ostrinia nubilalis larvae. PLoS One. 2012;7(8):e44090.
Yao, J., Buschman, L. L., Oppert, B., Khajuria, C., & Zhu, K. Y. (2012). Characterization of cDNAs encoding serine proteases and their transcriptional responses to Cry1Ab protoxin in the gut of Ostrinia nubilalis larvae. PloS One, 7(8), e44090. https://doi.org/10.1371/journal.pone.0044090
Yao J, et al. Characterization of cDNAs Encoding Serine Proteases and Their Transcriptional Responses to Cry1Ab Protoxin in the Gut of Ostrinia Nubilalis Larvae. PLoS One. 2012;7(8):e44090. PubMed PMID: 22952884.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of cDNAs encoding serine proteases and their transcriptional responses to Cry1Ab protoxin in the gut of Ostrinia nubilalis larvae. AU - Yao,Jianxiu, AU - Buschman,Lawrent L, AU - Oppert,Brenda, AU - Khajuria,Chitvan, AU - Zhu,Kun Yan, Y1 - 2012/08/31/ PY - 2012/07/02/received PY - 2012/08/01/accepted PY - 2012/9/7/entrez PY - 2012/9/7/pubmed PY - 2013/2/8/medline SP - e44090 EP - e44090 JF - PloS one JO - PLoS One VL - 7 IS - 8 N2 - Serine proteases, such as trypsin and chymotrypsin, are the primary digestive enzymes in lepidopteran larvae, and are also involved in Bacillus thuringiensis (Bt) protoxin activation and protoxin/toxin degradation. We isolated and sequenced 34 cDNAs putatively encoding trypsins, chymotrypsins and their homologs from the European corn borer (Ostrinia nubilalis) larval gut. Our analyses of the cDNA-deduced amino acid sequences indicated that 12 were putative trypsins, 12 were putative chymotrypsins, and the remaining 10 were trypsin and chymotrypsin homologs that lack one or more conserved residues of typical trypsins and chymotrypsins. Reverse transcription PCR analysis indicated that all genes were highly expressed in gut tissues, but one group of phylogenetically-related trypsin genes, OnTry-G2, was highly expressed in larval foregut and midgut, whereas another group, OnTry-G3, was highly expressed in the midgut and hindgut. Real-time quantitative PCR analysis indicated that several trypsin genes (OnTry5 and OnTry6) were significantly up-regulated in the gut of third-instar larvae after feeding on Cry1Ab protoxin from 2 to 24 h, whereas one trypsin (OnTry2) was down-regulated at all time points. Four chymotrypsin and chymotrypsin homolog genes (OnCTP2, OnCTP5, OnCTP12 and OnCTP13) were up-regulated at least 2-fold in the gut of the larvae after feeding on Cry1Ab protoxin for 24 h. Our data represent the first in-depth study of gut transcripts encoding expanded families of protease genes in O. nubilalis larvae and demonstrate differential expression of protease genes that may be related to Cry1Ab intoxication and/or resistance. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/22952884/Characterization_of_cDNAs_encoding_serine_proteases_and_their_transcriptional_responses_to_Cry1Ab_protoxin_in_the_gut_of_Ostrinia_nubilalis_larvae_ L2 - https://dx.plos.org/10.1371/journal.pone.0044090 DB - PRIME DP - Unbound Medicine ER -