Isolation and characterization of a cDNA encoding for a lysozyme from the gut of the reduviid bug <i>Triatoma infestans</i>

Kollien, Astrid H.; Fechner, Sabine; Waniek, Peter J.; Schaub, Günter A.

doi:10.1002/arch.10090

Cited by 55 publications

(78 citation statements)

References 38 publications

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“…The passage of the blood from the stomach to the digesting small intestine causes considerable destruction of symbiont populations, and only about 0,01% of the total population is still present in the rectum. 3 Since this development can not be correlated to the activity of lysozyme we are investigating other antibacterial compounds, e.g., defensins.…”

Section: Ve9 -Isolation and Characterization Of Defensin An Antimicrmentioning

confidence: 99%

“…The expression pattern of the lysozyme gene in the digestive tract of the bugs at different molting and feeding stages showed that this gene was upregulated directly after the molt and after feeding. 3 Investigating the expression of lysozymes in all parts used for activity tests via PCR amplification with specific lysozyme-primers, we obtained DNA-fragments by using cDNA of cardia, stomach, the final part of the small intestine and the salivary glands D2 and D3. Sequencing indicated that different lysozymes are expressed in the gut and salivary glands.…”

Section: Ve11 -Lysozymes Of Triatoma Infestansmentioning

confidence: 99%

“…Larvae placed for 24 h in suspensions of chitin azure were able to digest the ingested chitin. Semi-denaturing PAGE technique using glycol chitin and two fluorogenic substrate analogues showed the presence of three distinct chitinase activities: an endochitinase that catalyzed the hydrolysis of chitin; an endochitinase that cleaved the short substrate [4MU(GlcNAc) 3 ], and an exochitinase that hydrolyzed the chitobioside [4MU(GlcNAc) 2 ]. The endochitinase had an extremely broad pH-activity against glycol chitin and chitin azure, ranging from pH 4.0 to 10.0.…”

Section: Ve49 -Presence Of Chitinase Activities In the Gut Of Aedes Amentioning

confidence: 99%

“…The endochitinase had an extremely broad pH-activity against glycol chitin and chitin azure, ranging from pH 4.0 to 10.0. When the substrate [4MU(GlcNAc) 3 ] was used, two activities were observed at pH ranging from 4.0 to 6.0 and 8.0 to 10.0. Chitinase activity against [4MU(GlcNAc) 3 ] was detected throughout the gut, displaying the highest specific activity in the hindgut.…”

Section: Ve49 -Presence Of Chitinase Activities In the Gut Of Aedes Amentioning

confidence: 99%

“…When the substrate [4MU(GlcNAc) 3 ] was used, two activities were observed at pH ranging from 4.0 to 6.0 and 8.0 to 10.0. Chitinase activity against [4MU(GlcNAc) 3 ] was detected throughout the gut, displaying the highest specific activity in the hindgut. The pH values of the gut content were determined with the color shift indicators after larvae feeding.…”

Section: Ve49 -Presence Of Chitinase Activities In the Gut Of Aedes Amentioning

confidence: 99%

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Vectors

2003

Rev. Inst. Med. trop. S. Paulo

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Chagas disease is the most important parasitic disease in Latin America, as in Mexico, where 1.5-2% of blood donations are contaminated with antiTrypanosoma cruzi antibody. Through environmental and population based stratification, we estimate that 91 million inhabitants are at risk (78% through residence), 1,768,376 individuals are infected, and mortality may oscillate between 25,500 and 63,000 individuals/yr (830 of these are under 5 yrs old). The disease incidence is estimated at 69,000 cases/yr and approximately 530,500 individuals are currently in chronic phase. More than 96% of the transmission occurs via the vector, and niche modeling with GARP estimates that 67% of the transmission occurs via one of the 6 primary phyllosoma complex species.The economic loss due to incapacity is estimated at US$ 3,160,000,000/yr, while diagnostic and treatment costs currently could attain US$ 126,000,000/ yr. In the absence of a vector control program, disease burden could duplicate in 25 yrs, while chronic case treatment will augment by a factor of 45 over the same period. VE2 -MORPHOMETRIC ANALYSIS OF MEXICAN AND GUATEMALAN POPULATIONS OF TRIATOMA DIMIDIATA.Ramsey J.M., Lehmann P., Monroy C., Bargues M.D., Ordoñez R.Center for Infectious Disease Research, National Institute for Public Health, Cuernavaca, Morelos, Mexico Current estimates suggest that 2% of the Mexican population is seropositive for Trypanosoma cruzi, and the incidence of Chagas disease may supercede 69,000 cases/yr, with mortality equally 5-6% (25,000 cases) of the country´s total burden. Among the 18 vector species found in the country, Triatoma dimidiata plays a critical role in transmission principally in Gulf coast and all states south of the Istmus of Tehuantepec. Current molecular studies using ITS-2 rDNA and mitochondrial DNA markers, suggest that the populations found in the Yucatan peninsula may be an ancestral stock, from which other Mexican, Mesomerican, and Colombian/Venezuelan populations derived. Given profound genetic differences between yucatecan and all other populations, phenotypic differences might be expected using head and wing morphometric and symmetry discriminate analysis. 187 specimens of T. dimidiata collected from the states of Yucatán, Veracruz, and San Luis Potosí (SLP), were compared to a population from the Petén, Guatemala, and T. phyllosoma from Oaxaca as outgroup (:33 specimens per species). 12 head and 14 wing characters were measured separately for male and female samples, and measurements analyzed using multigroup principal component and discriminate analyses, sexual dimorphism and wing asymmetry. Guillaumin profiles indicated no size difference between SLP and Veracruz populations, while Peten specimens were smaller, and those from the Yucatan the smallest. None of the populations were separated completely using principal component analysis, although there was a clear tendency for separation. No shape differences were observed between SLP and Veracruz populations although they were clearly separated from both the Yu...

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Section: Ve9 -Isolation and Characterization Of Defensin An Antimicrmentioning

confidence: 99%

Section: Ve11 -Lysozymes Of Triatoma Infestansmentioning

confidence: 99%

Section: Ve49 -Presence Of Chitinase Activities In the Gut Of Aedes Amentioning

confidence: 99%

Section: Ve49 -Presence Of Chitinase Activities In the Gut Of Aedes Amentioning

confidence: 99%

Section: Ve49 -Presence Of Chitinase Activities In the Gut Of Aedes Amentioning

confidence: 99%

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Vectors

2003

Rev. Inst. Med. trop. S. Paulo

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Up‐regulation of lysozyme gene expression during metamorphosis and immune challenge of the cotton bollworm, Helicoverpa armigera

Zhang

Huang

Zhou

et al. 2008

Arch Insect Biochem Physiol

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Lysozymes act as crucial bacteriolytic enzymes in insect immune system by hydrolyzing the beta (1-->4) bonds between N-acetylglucosamine and N-acetylmuramic acid in the peptidoglycan of prokaryotic cell walls. We have isolated and characterized a Helicoverpa armigera cDNA encoding an insect lysozyme named HaLyz. We amplified a fragment by PCR, using degenerate primers derived from the conservative amino acid sequences for performing 5' and 3' RACE. The full-length cDNA was 661 base pairs. The theoretical pI and molecular weight of the protein were computed to be 9.08 and 15.6 kDa, respectively. Prokaryotic expression of the HaLyz ORF by Escherichia coli confirmed the calculated molecular weight of the protein. The deduced 135 amino acids showed high homology with known lysozymes from other insects, ranging from 47% to 89% by BLASTp search in NCBI. Analyses revealed that this protein has a typical lysozyme C signature among amino acids 93-111, CNVTCAEMLLDDITKASTC. An interesting relation between immunity and larva to pupa metamorphosis in insects was discovered. Real time-PCR showed that HaLyz gene expression was transiently enhanced at the onset of metamorphosis of the cotton bollworm, Helicoverpa armigera. The gene expression was up-regulated after the injection of E. coli or entomopathogenic fungi, Beauveria bassiana, but showed different expression patterns.

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Bacteriolytic activity in saliva of the hematophagous Triatoma infestans (Reduviidae) and novel characterization and expression site of a third lysozyme

Meiser

Klenner

Balczun

et al. 2023

Arch Insect Biochem Physiol

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Saliva of hematophagous insects contains many different compounds, mainly acting as anticoagulants. Investigating the bacteriolytic compounds of the saliva of the bloodsucking Triatoma infestans photometrically between pH 3 and pH 10 using unfed fifth instars and nymphs up to 15 days after feeding, we found bacteriolytic activity against lyophilized Micrococcus luteus was stronger at pH 4 and pH 6. After feeding, the activity level at pH 4 was unchanged, but at pH 6 more than doubled between 3 and 7 days after feeding. In zymographs of the saliva and after incubation at pH 4, bacteriolytic activity against Micrococcus luteus was present at eight lysis zones between 14.1 and 38.5 kDa, showing the strongest activity at 24.5 kDa. After incubation at pH 6, lysis zones only appeared at 15.3, 17, and 31.4 kDa. Comparing zymographs of the saliva of unfed and fed nymphs, bacteriolytic activity at 17 kDa increased after feeding. In total nine lysis bands appeared, also at >30 kDa, so far unreported in the saliva of triatomines. Reverse transcription polymerase chain reaction using oligonucleotides based on the previously described lysozyme gene of T. infestans, TiLys1, verified expression of genes encoding TiLys1 and TiLys2 in the salivary glands, but also of an undescribed third lysozyme, TiLys3, of which the cloned cDNA shares characteristics with other c‐type lysozymes of insects. While TiLys1 was expressed in the tissue of all three salivary glands, transcripts of TiLys2 and of TiLys3 seem to be present only in the gland G1 and G3, respectively.

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Isolation and characterization of a cDNA encoding for a lysozyme from the gut of the reduviid bug Triatoma infestans

Cited by 55 publications

References 38 publications

Vectors

Vectors

Up‐regulation of lysozyme gene expression during metamorphosis and immune challenge of the cotton bollworm, Helicoverpa armigera

Bacteriolytic activity in saliva of the hematophagous Triatoma infestans (Reduviidae) and novel characterization and expression site of a third lysozyme

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