ABSTRACT
Several receptor proteins of Cry toxin have been previously identified, including cadherin-like, aminopeptidase N, and alkaline phosphatase. In the present work, a novel binding protein, V-ATPase subunit A (HpVAA), was identified in Holotricia parallela larvae and characterized. We performed reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends technology to obtain the cDNA of the full-length hpvaa. Sequencing analysis showed that the open reading frame of hpvaa (GenBank accession No. KU497557) is 1845 bp long, encoding 614 amino acid residues. The predicted molecular weight and isoelectric point of HpVAA were 67.85 kDa and 4.9, respectively. The HpVAA protein, which includes two putative conserved domains, ATP-synt_ab_N and ATP-synt_ab_C, and a Walker A (GAFGCGKT) motif and a Walker B (SMMAD) motif, possesses the same structural characteristics as V-ATPase subunit A from other insects. The protein was successfully expressed in Escherichia coli, and a ligand blot assay showed binding of the protein with Cry8Ea3 toxin. Transcriptional analysis of hpvaa in different tissues of H. parallela larvae was performed by qRT-PCR, which showed that the relative expression of hpvaa in the Malpighian tubules is higher than that in other tissues.
Subject(s)
Bacterial Proteins/chemistry , Coleoptera/genetics , Endotoxins/chemistry , Hemolysin Proteins/chemistry , Insect Proteins/chemistry , Phylogeny , Protein Subunits/chemistry , Vacuolar Proton-Translocating ATPases/chemistry , Amino Acid Sequence , Animals , Bacillus thuringiensis Toxins , Bacterial Proteins/genetics , Base Sequence , Cloning, Molecular , Coleoptera/classification , Coleoptera/immunology , Coleoptera/microbiology , Conserved Sequence , DNA, Complementary/genetics , DNA, Complementary/metabolism , Endotoxins/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Hemolysin Proteins/genetics , Insect Proteins/genetics , Insect Proteins/immunology , Isoelectric Point , Larva/genetics , Larva/microbiology , Malpighian Tubules/immunology , Malpighian Tubules/metabolism , Malpighian Tubules/microbiology , Models, Molecular , Molecular Weight , Open Reading Frames , Protein Binding , Protein Subunits/genetics , Protein Subunits/immunology , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Vacuolar Proton-Translocating ATPases/genetics , Vacuolar Proton-Translocating ATPases/immunologyABSTRACT
Given the physiological importance of the Malpighian tubules to homeostasis in ants, this study aimed to characterize the enzymology, histology, histochemistry, and ultramorphology of the Malpighian tubules of Cephalotes atratus, C. clypeatus, and C. pusillus, as a contribution for the understanding of this organ, as well as to examine its role in the maintenance of symbiontic microorganisms in the ileum of these ants.
Subject(s)
Ants/microbiology , Malpighian Tubules/microbiology , Symbiosis , Adenosine Triphosphatases/analysis , Animals , Hydrogen-Ion Concentration , Malpighian Tubules/enzymology , Malpighian Tubules/ultrastructureABSTRACT
There are very few reports on the microbiota of the digestive tract of sand flies, an important omission considering that blood is not the only meal ingested. Male and female sand flies obtain sugar meals from several sources, thereby increasing their chance of infection with microorganisms. Chances of contamination are higher when insects are bred in the laboratory, and this may affect the development of Leishmania spp. From the digestive tract of 300 sand fly females separated in two groups we isolated 10 species of bacteria in group 1 and 8 species in group 2. In group 1, Enterobacteriaceae of the following genera were identified: Serratia, Enterobacter, and Yokenella and the non-fermenters: Pseudomonas, Acinetobacter, and Stenotrophomonas. In group 2, the Enterobacteriaceae Enterobacter and Serratia were identified as well as the non-fermenters Acinetobacter, Stenotrophomonas, Burkolderia, and Pseudomonas.
Subject(s)
Digestive System/microbiology , Malpighian Tubules/microbiology , Psychodidae/microbiology , Animals , Blood , Carbohydrates , Female , FoodABSTRACT
No presente trabalho foram dissecados o trato digestivo de 245 fêmeas de Lutzomyia longipalpis originários da Gruta da Lapinha, Município de Lagoa Santa, MG, formando 7 grupos de 35 flebotomíneos. Das 8 espécies de bactérias isoladas houve uma predominância de bactérias Gram negativas (BGN) pertencentes ao grupo de näo fermentadoras de açúcar das seguintes espécies: Acinetobacter lowffii, Stenotrophomonas maltophhilia, Pseudomonas putida e Flavimonas orizihabitans. No grupo das fermentadoras tivemos: Enterobacter cloacae e Klebsiella ozaenae. No grupo dos Gram positivos foram identificados Bacillus thuringiensis e Staphylococcus spp