Molecular identification, characterization, and expression analysis of a serine protease inhibitor gene from cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae) / Identificação molecular, caracterização e análise de expressão de um gene inibidor de serina protease da lagarta Helicoverpa armigera (Lepidoptera: noctuidae) na cultura de algodão

Serine protease inhibitors (serpins), a superfamily of protease inhibitors, are known to be involved in several physiological processes, such as development, metamorphosis, and innate immunity. In our study, a full-length serpin cDNA, designated Haserpin1, was isolated from the cotton bollworm Helicoverpa armigera. The cDNA sequence of Haserpin1 is 1176 nt long, with an open reading frame encoding 391 amino acids; there is one exon and no intron. The predicted molecular weight of Haserpin1 is 43.53 kDa, with an isoelectric point of 4.98. InterProScan was employed for Haserpin1 functional characterization, which revealed that Haserpin1 contains highly conserved signature motifs, including a reactive center loop (RCL) with a hinge region (E341N350), the serpin signature, (F367F375) and a predicted P1P1 cleavage site (L357S358), which are useful for identifying serpins. Transcripts of Haserpin1 were constitutively expressed in the fat body, suggesting that it is the major site for serpin synthesis. During the developmental stages, a fluctuation in the expression level of Haserpin1 was observed, with low expression detected at the 5th-instar larval stage. In contrast, relatively high expression was detected at the prepupal stage, suggesting that Haserpin1 might play a critical role at the H. armigera wandering stage. Although the detailed function of this serpin (Haserpin1) needs to be elucidated, our study provides a perspective for the functional investigation of serine protease inhibitor genes.(AU)

Sabe-se que os inibidores de serina protease (serpinas), uma superfamília de inibidores de protease, estão envolvidos em vários processos fisiológicos, como desenvolvimento, metamorfose e imunidade inata. Neste estudo, um cDNA de serpina de comprimento total, denominado Haserpin1, foi isolado da lagarta Helicoverpa armigera na cultura de algodão. A sequência de ADNc de Haserpin1 tem 1.176 nt de comprimento, com uma grelha de leitura aberta que codifica 391 aminoácidos; existe um éxon, mas nenhum íntron. O peso molecular previsto de Haserpin1 é de 43,53 kDa, com um ponto isoelétrico de 4,98. O InterProScan foi empregado para a caracterização funcional do Haserpin1, que revelou que o Haserpin1 contém motivos de assinatura altamente conservados, incluindo um loop central reativo (RCL) com uma região de dobradiça (E341-N350), a assinatura da serpina (F367-F375) e um local de clivagem previsto de P1-P1' (L357-S358), que são úteis para identificar serpinas. As transcrições de Haserpin1 foram expressas constitutivamente no corpo gordo, sugerindo que é o principal local para a síntese de serpinas. Durante os estágios de desenvolvimento, observou-se uma flutuação no nível de expressão de Haserpin1, com baixa expressão detectada no estágio larval do 5º ínstar. Por outro lado, detectou-se uma expressão relativamente alta no estágio pré-pupal, sugerindo que o Haserpin1 pode desempenhar um papel crítico no estágio errante de H. armigera. Embora a função detalhada dessa serpina (Haserpin1) precise ser elucidada, este estudo fornece uma perspectiva para a investigação funcional dos genes inibidores da serina protease.(AU)

Molecular identification, characterization, and expression analysis of a serine protease inhibitor gene from cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

Abstract Serine protease inhibitors (serpins), a superfamily of protease inhibitors, are known to be involved in several physiological processes, such as development, metamorphosis, and innate immunity. In our study, a full-length serpin cDNA, designated Haserpin1, was isolated from the cotton bollworm Helicoverpa armigera. The cDNA sequence of Haserpin1 is 1176 nt long, with an open reading frame encoding 391 amino acids; there is one exon and no intron. The predicted molecular weight of Haserpin1 is 43.53 kDa, with an isoelectric point of 4.98. InterProScan was employed for Haserpin1 functional characterization, which revealed that Haserpin1 contains highly conserved signature motifs, including a reactive center loop (RCL) with a hinge region (E341N350), the serpin signature, (F367F375) and a predicted P1P1 cleavage site (L357S358), which are useful for identifying serpins. Transcripts of Haserpin1 were constitutively expressed in the fat body, suggesting that it is the major site for serpin synthesis. During the developmental stages, a fluctuation in the expression level of Haserpin1 was observed, with low expression detected at the 5th-instar larval stage. In contrast, relatively high expression was detected at the prepupal stage, suggesting that Haserpin1 might play a critical role at the H. armigera wandering stage. Although the detailed function of this serpin (Haserpin1) needs to be elucidated, our study provides a perspective for the functional investigation of serine protease inhibitor genes.

Resumo Sabe-se que os inibidores de serina protease (serpinas), uma superfamília de inibidores de protease, estão envolvidos em vários processos fisiológicos, como desenvolvimento, metamorfose e imunidade inata. Neste estudo, um cDNA de serpina de comprimento total, denominado Haserpin1, foi isolado da lagarta Helicoverpa armigera na cultura de algodão. A sequência de ADNc de Haserpin1 tem 1.176 nt de comprimento, com uma grelha de leitura aberta que codifica 391 aminoácidos; existe um éxon, mas nenhum íntron. O peso molecular previsto de Haserpin1 é de 43,53 kDa, com um ponto isoelétrico de 4,98. O InterProScan foi empregado para a caracterização funcional do Haserpin1, que revelou que o Haserpin1 contém motivos de assinatura altamente conservados, incluindo um loop central reativo (RCL) com uma região de dobradiça (E341-N350), a assinatura da serpina (F367-F375) e um local de clivagem previsto de P1-P1' (L357-S358), que são úteis para identificar serpinas. As transcrições de Haserpin1 foram expressas constitutivamente no corpo gordo, sugerindo que é o principal local para a síntese de serpinas. Durante os estágios de desenvolvimento, observou-se uma flutuação no nível de expressão de Haserpin1, com baixa expressão detectada no estágio larval do 5º ínstar. Por outro lado, detectou-se uma expressão relativamente alta no estágio pré-pupal, sugerindo que o Haserpin1 pode desempenhar um papel crítico no estágio errante de H. armigera. Embora a função detalhada dessa serpina (Haserpin1) precise ser elucidada, este estudo fornece uma perspectiva para a investigação funcional dos genes inibidores da serina protease.

Molecular identification, characterization, and expression analysis of a serine protease inhibitor gene from cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

Abstract Serine protease inhibitors (serpins), a superfamily of protease inhibitors, are known to be involved in several physiological processes, such as development, metamorphosis, and innate immunity. In our study, a full-length serpin cDNA, designated Haserpin1, was isolated from the cotton bollworm Helicoverpa armigera. The cDNA sequence of Haserpin1 is 1176 nt long, with an open reading frame encoding 391 amino acids; there is one exon and no intron. The predicted molecular weight of Haserpin1 is 43.53 kDa, with an isoelectric point of 4.98. InterProScan was employed for Haserpin1 functional characterization, which revealed that Haserpin1 contains highly conserved signature motifs, including a reactive center loop (RCL) with a hinge region (E341N350), the serpin signature, (F367F375) and a predicted P1P1 cleavage site (L357S358), which are useful for identifying serpins. Transcripts of Haserpin1 were constitutively expressed in the fat body, suggesting that it is the major site for serpin synthesis. During the developmental stages, a fluctuation in the expression level of Haserpin1 was observed, with low expression detected at the 5th-instar larval stage. In contrast, relatively high expression was detected at the prepupal stage, suggesting that Haserpin1 might play a critical role at the H. armigera wandering stage. Although the detailed function of this serpin (Haserpin1) needs to be elucidated, our study provides a perspective for the functional investigation of serine protease inhibitor genes.

Resumo Sabe-se que os inibidores de serina protease (serpinas), uma superfamília de inibidores de protease, estão envolvidos em vários processos fisiológicos, como desenvolvimento, metamorfose e imunidade inata. Neste estudo, um cDNA de serpina de comprimento total, denominado Haserpin1, foi isolado da lagarta Helicoverpa armigera na cultura de algodão. A sequência de ADNc de Haserpin1 tem 1.176 nt de comprimento, com uma grelha de leitura aberta que codifica 391 aminoácidos; existe um éxon, mas nenhum íntron. O peso molecular previsto de Haserpin1 é de 43,53 kDa, com um ponto isoelétrico de 4,98. O InterProScan foi empregado para a caracterização funcional do Haserpin1, que revelou que o Haserpin1 contém motivos de assinatura altamente conservados, incluindo um loop central reativo (RCL) com uma região de dobradiça (E341-N350), a assinatura da serpina (F367-F375) e um local de clivagem previsto de P1-P1' (L357-S358), que são úteis para identificar serpinas. As transcrições de Haserpin1 foram expressas constitutivamente no corpo gordo, sugerindo que é o principal local para a síntese de serpinas. Durante os estágios de desenvolvimento, observou-se uma flutuação no nível de expressão de Haserpin1, com baixa expressão detectada no estágio larval do 5º ínstar. Por outro lado, detectou-se uma expressão relativamente alta no estágio pré-pupal, sugerindo que o Haserpin1 pode desempenhar um papel crítico no estágio errante de H. armigera. Embora a função detalhada dessa serpina (Haserpin1) precise ser elucidada, este estudo fornece uma perspectiva para a investigação funcional dos genes inibidores da serina protease.

Recent topics of candidate antigens for immunological control of ixodid ticks

Ticks have been recognized as harmful parasite since ancient times. At present immunological protection of host against ticks is the most practical and sustainable tick control method, which is more suitable to natural environment compared to the current use of acaricides. Recently, focuses on the development of anti-tick vaccine are the identification, molecular cloning and in vitro production of recombinant protein, responsible for executing key roles in regulating physiology, modulation of host immune response and pathogen transmission via ticks. Among several works, serine protease inhibitors have been thought as one of the most interest vaccine candidates, because serine protease inhibitors are mainly involved in the maintenance of homeostasis. In the current review, we would like to introduce selected examples covering aspects of tick vaccine antigen identification and analyses, because advances in vector molecular biology open new possibilities for vaccine development. In dealing with this subject, contents were mainly divided into tick salivary gland associated molecules (exposed antigens) injected into the host during tick feeding and no salivary gland molecules (concealed antigens). While emerging the fact that serine protease inhibitors belong to either exposed or concealed antigens, the utility of serine protease inhibitors for the candidate vaccine have been discus

Recent topics of candidate antigens for immunological control of ixodid ticks

Ticks have been recognized as harmful parasite since ancient times. At present immunological protection of host against ticks is the most practical and sustainable tick control method, which is more suitable to natural environment compared to the current use of acaricides. Recently, focuses on the development of anti-tick vaccine are the identification, molecular cloning and in vitro production of recombinant protein, responsible for executing key roles in regulating physiology, modulation of host immune response and pathogen transmission via ticks. Among several works, serine protease inhibitors have been thought as one of the most interest vaccine candidates, because serine protease inhibitors are mainly involved in the maintenance of homeostasis. In the current review, we would like to introduce selected examples covering aspects of tick vaccine antigen identification and analyses, because advances in vector molecular biology open new possibilities for vaccine development. In dealing with this subject, contents were mainly divided into tick salivary gland associated molecules (exposed antigens) injected into the host during tick feeding and no salivary gland molecules (concealed antigens). While emerging the fact that serine protease inhibitors belong to either exposed or concealed antigens, the utility of serine protease inhibitors for the candidate vaccine have been discus

Recent topics of candidate antigens for immunological control of ixodid ticks

Ticks have been recognized as harmful parasite since ancient times. At present immunological protection of host against ticks is the most practical and sustainable tick control method, which is more suitable to natural environment compared to the current use of acaricides. Recently, focuses on the development of anti-tick vaccine are the identification, molecular cloning and in vitro production of recombinant protein, responsible for executing key roles in regulating physiology, modulation of host immune response and pathogen transmission via ticks. Among several works, serine protease inhibitors have been thought as one of the most interest vaccine candidates, because serine protease inhibitors are mainly involved in the maintenance of homeostasis. In the current review, we would like to introduce selected examples covering aspects of tick vaccine antigen identification and analyses, because advances in vector molecular biology open new possibilities for vaccine development. In dealing with this subject, contents were mainly divided into tick salivary gland associated molecules (exposed antigens) injected into the host during tick feeding and no salivary gland molecules (concealed antigens). While emerging the fact that serine protease inhibitors belong to either exposed or concealed antigens, the utility of serine protease inhibitors for the candidate vaccine have been discus

Recent topics of candidate antigens for immunological control of ixodid ticks

Ticks have been recognized as harmful parasite since ancient times. At present immunological protection of host against ticks is the most practical and sustainable tick control method, which is more suitable to natural environment compared to the current use of acaricides. Recently, focuses on the development of anti-tick vaccine are the identification, molecular cloning and in vitro production of recombinant protein, responsible for executing key roles in regulating physiology, modulation of host immune response and pathogen transmission via ticks. Among several works, serine protease inhibitors have been thought as one of the most interest vaccine candidates, because serine protease inhibitors are mainly involved in the maintenance of homeostasis. In the current review, we would like to introduce selected examples covering aspects of tick vaccine antigen identification and analyses, because advances in vector molecular biology open new possibilities for vaccine development. In dealing with this subject, contents were mainly divided into tick salivary gland associated molecules (exposed antigens) injected into the host during tick feeding and no salivary gland molecules (concealed antigens). While emerging the fact that serine protease inhibitors belong to either exposed or concealed antigens, the utility of serine protease inhibitors for the candidate vaccine have been discus

Recent topics of candidate antigens for immunological control of ixodid ticks

Ticks have been recognized as harmful parasite since ancient times. At present immunological protection of host against ticks is the most practical and sustainable tick control method, which is more suitable to natural environment compared to the current use of acaricides. Recently, focuses on the development of anti-tick vaccine are the identification, molecular cloning and in vitro production of recombinant protein, responsible for executing key roles in regulating physiology, modulation of host immune response and pathogen transmission via ticks. Among several works, serine protease inhibitors have been thought as one of the most interest vaccine candidates, because serine protease inhibitors are mainly involved in the maintenance of homeostasis. In the current review, we would like to introduce selected examples covering aspects of tick vaccine antigen identification and analyses, because advances in vector molecular biology open new possibilities for vaccine development. In dealing with this subject, contents were mainly divided into tick salivary gland associated molecules (exposed antigens) injected into the host during tick feeding and no salivary gland molecules (concealed antigens). While emerging the fact that serine protease inhibitors belong to either exposed or concealed antigens, the utility of serine protease inhibitors for the candidate vaccine have been discus

Recent topics of candidate antigens for immunological control of ixodid ticks

Ticks have been recognized as harmful parasite since ancient times. At present immunological protection of host against ticks is the most practical and sustainable tick control method, which is more suitable to natural environment compared to the current use of acaricides. Recently, focuses on the development of anti-tick vaccine are the identification, molecular cloning and in vitro production of recombinant protein, responsible for executing key roles in regulating physiology, modulation of host immune response and pathogen transmission via ticks. Among several works, serine protease inhibitors have been thought as one of the most interest vaccine candidates, because serine protease inhibitors are mainly involved in the maintenance of homeostasis. In the current review, we would like to introduce selected examples covering aspects of tick vaccine antigen identification and analyses, because advances in vector molecular biology open new possibilities for vaccine development. In dealing with this subject, contents were mainly divided into tick salivary gland associated molecules (exposed antigens) injected into the host during tick feeding and no salivary gland molecules (concealed antigens). While emerging the fact that serine protease inhibitors belong to either exposed or concealed antigens, the utility of serine protease inhibitors for the candidate vaccine have been discus