Molecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pests
Main Author: | |
---|---|
Publication Date: | 2005 |
Other Authors: | , , , , , , |
Format: | Article |
Language: | eng |
Source: | Repositório Institucional da UCB |
Download full: | http://twingo.ucb.br:8080/jspui/handle/10869/640 https://repositorio.ucb.br:9443/jspui/handle/123456789/7807 |
Summary: | Alpha-amylase inhibitors have important roles in plant defense mechanisms, particularly against insects, and several of these inhibitors have been expressed in different crops to increase their resistance to particular insects. In this work, we report the cloning and expression of a gene encoding for a new a-amylase inhibitor (BIII) from rye (Secale cereale) seeds. The BIII gene contains 354 nucleotides that encode for 118 amino acids sequence. A 313 bp fragment of the gene was expressed in Escherichia coli and resulted in a functional inhibitor that reduced the activity of a-amylases of larvae of the coleopteran pests Acanthoscelides obtectus, Zabrotess subfasciatus and Anthonomus grandis. In contrast, the inhibitor did not inhibit the activity of porcine pancreatic a-amylase. Although the amino acid sequence of BIII showed high identity with those of bifunctional inhibitors, the recombinant protein was unable to inhibit trypsin-like serine proteinases. The effects of recombinant BIII were evaluated in vivo against A. grandis. When first instar larvae were reared on an artificial diet containing four different concentrations of BIII, a reduction in larval weight and a mortality of 83% were observed at the highest concentration. |
id |
UCB-2_73cb5c9ddf58b48d5dc30a0cc868f670 |
---|---|
oai_identifier_str |
oai:200.214.135.189:123456789/7807 |
network_acronym_str |
UCB-2 |
network_name_str |
Repositório Institucional da UCB |
spelling |
Dias, Simoni C.Franco, Octávio L.Magalhães, Cláudio P.Oliveira-Neto, Osmundo Brilhante deLaumann, Raúl A.Figueira, Edson L. Z.Melo, Francislete R.Grossi-de-Sá, Maria F.2016-10-10T03:52:44Z2016-10-10T03:52:44Z2005-02DIAS, Simoni C. et al. Molecular Cloning and Expression of an a-Amylase Inhibitor. The Protein Journal, v. 24, n.. 2, p. 113-123, 2005.http://twingo.ucb.br:8080/jspui/handle/10869/640https://repositorio.ucb.br:9443/jspui/handle/123456789/7807Alpha-amylase inhibitors have important roles in plant defense mechanisms, particularly against insects, and several of these inhibitors have been expressed in different crops to increase their resistance to particular insects. In this work, we report the cloning and expression of a gene encoding for a new a-amylase inhibitor (BIII) from rye (Secale cereale) seeds. The BIII gene contains 354 nucleotides that encode for 118 amino acids sequence. A 313 bp fragment of the gene was expressed in Escherichia coli and resulted in a functional inhibitor that reduced the activity of a-amylases of larvae of the coleopteran pests Acanthoscelides obtectus, Zabrotess subfasciatus and Anthonomus grandis. In contrast, the inhibitor did not inhibit the activity of porcine pancreatic a-amylase. Although the amino acid sequence of BIII showed high identity with those of bifunctional inhibitors, the recombinant protein was unable to inhibit trypsin-like serine proteinases. The effects of recombinant BIII were evaluated in vivo against A. grandis. When first instar larvae were reared on an artificial diet containing four different concentrations of BIII, a reduction in larval weight and a mortality of 83% were observed at the highest concentration.Made available in DSpace on 2016-10-10T03:52:44Z (GMT). No. of bitstreams: 5 Molecular Cloning and Expression.PDF: 661224 bytes, checksum: 77f2050b845e88723a63c4164c1a0de9 (MD5) license_url: 52 bytes, checksum: 2f32edb9c19a57e928372a33fd08dba5 (MD5) license_text: 24372 bytes, checksum: 94b0a37ff5ec51de8c55507bff4a7ff9 (MD5) license_rdf: 24623 bytes, checksum: 378d22d8fe50e084ee2f354be78cbe62 (MD5) license.txt: 1887 bytes, checksum: 445d1980f282ec865917de35a4c622f6 (MD5) Previous issue date: 2005-02SimPublicadoTextoA-Amylase InhibitorCM protein, insect pestMolecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pestsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleThe Protein Journalinfo:eu-repo/semantics/openAccessengreponame:Repositório Institucional da UCBinstname:Universidade Católica de Brasília (UCB)instacron:UCBORIGINALMolecular Cloning and Expression.PDFapplication/pdf661224https://200.214.135.178:9443/jspui/bitstream/123456789/7807/1/Molecular%20Cloning%20and%20Expression.PDF77f2050b845e88723a63c4164c1a0de9MD51CC-LICENSElicense_urlapplication/octet-stream52https://200.214.135.178:9443/jspui/bitstream/123456789/7807/2/license_url2f32edb9c19a57e928372a33fd08dba5MD52license_textapplication/octet-stream24372https://200.214.135.178:9443/jspui/bitstream/123456789/7807/3/license_text94b0a37ff5ec51de8c55507bff4a7ff9MD53license_rdfapplication/octet-stream24623https://200.214.135.178:9443/jspui/bitstream/123456789/7807/4/license_rdf378d22d8fe50e084ee2f354be78cbe62MD54LICENSElicense.txttext/plain1887https://200.214.135.178:9443/jspui/bitstream/123456789/7807/5/license.txt445d1980f282ec865917de35a4c622f6MD55TEXTMolecular Cloning and Expression.PDF.txtMolecular Cloning and Expression.PDF.txtExtracted texttext/plain40943https://200.214.135.178:9443/jspui/bitstream/123456789/7807/6/Molecular%20Cloning%20and%20Expression.PDF.txt2bc9ccb7ea5cddeb66985787fce9b042MD56123456789/78072017-01-17 15:11:00.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ório de Publicaçõeshttps://repositorio.ucb.br:9443/jspui/ |
dc.title.pt_BR.fl_str_mv |
Molecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pests |
title |
Molecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pests |
spellingShingle |
Molecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pests Dias, Simoni C. A-Amylase Inhibitor CM protein, insect pest |
title_short |
Molecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pests |
title_full |
Molecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pests |
title_fullStr |
Molecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pests |
title_full_unstemmed |
Molecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pests |
title_sort |
Molecular Cloning and Expression of an a-Amylase Inhibitor from Rye with Potential for Controlling Insect Pests |
author |
Dias, Simoni C. |
author_facet |
Dias, Simoni C. Franco, Octávio L. Magalhães, Cláudio P. Oliveira-Neto, Osmundo Brilhante de Laumann, Raúl A. Figueira, Edson L. Z. Melo, Francislete R. Grossi-de-Sá, Maria F. |
author_role |
author |
author2 |
Franco, Octávio L. Magalhães, Cláudio P. Oliveira-Neto, Osmundo Brilhante de Laumann, Raúl A. Figueira, Edson L. Z. Melo, Francislete R. Grossi-de-Sá, Maria F. |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Dias, Simoni C. Franco, Octávio L. Magalhães, Cláudio P. Oliveira-Neto, Osmundo Brilhante de Laumann, Raúl A. Figueira, Edson L. Z. Melo, Francislete R. Grossi-de-Sá, Maria F. |
dc.subject.por.fl_str_mv |
A-Amylase Inhibitor CM protein, insect pest |
topic |
A-Amylase Inhibitor CM protein, insect pest |
dc.description.abstract.por.fl_txt_mv |
Alpha-amylase inhibitors have important roles in plant defense mechanisms, particularly against insects, and several of these inhibitors have been expressed in different crops to increase their resistance to particular insects. In this work, we report the cloning and expression of a gene encoding for a new a-amylase inhibitor (BIII) from rye (Secale cereale) seeds. The BIII gene contains 354 nucleotides that encode for 118 amino acids sequence. A 313 bp fragment of the gene was expressed in Escherichia coli and resulted in a functional inhibitor that reduced the activity of a-amylases of larvae of the coleopteran pests Acanthoscelides obtectus, Zabrotess subfasciatus and Anthonomus grandis. In contrast, the inhibitor did not inhibit the activity of porcine pancreatic a-amylase. Although the amino acid sequence of BIII showed high identity with those of bifunctional inhibitors, the recombinant protein was unable to inhibit trypsin-like serine proteinases. The effects of recombinant BIII were evaluated in vivo against A. grandis. When first instar larvae were reared on an artificial diet containing four different concentrations of BIII, a reduction in larval weight and a mortality of 83% were observed at the highest concentration. |
dc.description.version.pt_BR.fl_txt_mv |
Sim |
dc.description.status.pt_BR.fl_txt_mv |
Publicado |
description |
Alpha-amylase inhibitors have important roles in plant defense mechanisms, particularly against insects, and several of these inhibitors have been expressed in different crops to increase their resistance to particular insects. In this work, we report the cloning and expression of a gene encoding for a new a-amylase inhibitor (BIII) from rye (Secale cereale) seeds. The BIII gene contains 354 nucleotides that encode for 118 amino acids sequence. A 313 bp fragment of the gene was expressed in Escherichia coli and resulted in a functional inhibitor that reduced the activity of a-amylases of larvae of the coleopteran pests Acanthoscelides obtectus, Zabrotess subfasciatus and Anthonomus grandis. In contrast, the inhibitor did not inhibit the activity of porcine pancreatic a-amylase. Although the amino acid sequence of BIII showed high identity with those of bifunctional inhibitors, the recombinant protein was unable to inhibit trypsin-like serine proteinases. The effects of recombinant BIII were evaluated in vivo against A. grandis. When first instar larvae were reared on an artificial diet containing four different concentrations of BIII, a reduction in larval weight and a mortality of 83% were observed at the highest concentration. |
publishDate |
2005 |
dc.date.issued.fl_str_mv |
2005-02 |
dc.date.accessioned.fl_str_mv |
2016-10-10T03:52:44Z |
dc.date.available.fl_str_mv |
2016-10-10T03:52:44Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
status_str |
publishedVersion |
format |
article |
dc.identifier.citation.fl_str_mv |
DIAS, Simoni C. et al. Molecular Cloning and Expression of an a-Amylase Inhibitor. The Protein Journal, v. 24, n.. 2, p. 113-123, 2005. |
dc.identifier.uri.fl_str_mv |
http://twingo.ucb.br:8080/jspui/handle/10869/640 https://repositorio.ucb.br:9443/jspui/handle/123456789/7807 |
identifier_str_mv |
DIAS, Simoni C. et al. Molecular Cloning and Expression of an a-Amylase Inhibitor. The Protein Journal, v. 24, n.. 2, p. 113-123, 2005. |
url |
http://twingo.ucb.br:8080/jspui/handle/10869/640 https://repositorio.ucb.br:9443/jspui/handle/123456789/7807 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
Texto |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UCB instname:Universidade Católica de Brasília (UCB) instacron:UCB |
instname_str |
Universidade Católica de Brasília (UCB) |
instacron_str |
UCB |
institution |
UCB |
reponame_str |
Repositório Institucional da UCB |
collection |
Repositório Institucional da UCB |
bitstream.url.fl_str_mv |
https://200.214.135.178:9443/jspui/bitstream/123456789/7807/1/Molecular%20Cloning%20and%20Expression.PDF https://200.214.135.178:9443/jspui/bitstream/123456789/7807/2/license_url https://200.214.135.178:9443/jspui/bitstream/123456789/7807/3/license_text https://200.214.135.178:9443/jspui/bitstream/123456789/7807/4/license_rdf https://200.214.135.178:9443/jspui/bitstream/123456789/7807/5/license.txt https://200.214.135.178:9443/jspui/bitstream/123456789/7807/6/Molecular%20Cloning%20and%20Expression.PDF.txt |
bitstream.checksum.fl_str_mv |
77f2050b845e88723a63c4164c1a0de9 2f32edb9c19a57e928372a33fd08dba5 94b0a37ff5ec51de8c55507bff4a7ff9 378d22d8fe50e084ee2f354be78cbe62 445d1980f282ec865917de35a4c622f6 2bc9ccb7ea5cddeb66985787fce9b042 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
|
repository.mail.fl_str_mv |
|
_version_ |
1724829831361724416 |