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Development of genetic system to inactivate a Borrelia turicatae surface protein selectively produced within the salivary glands of the arthropod vector.

Lopez JE, Wilder HK, Hargrove R, Brooks CP, Peterson KE, Beare PA, Sturdevant DE, Nagarajan V, Raffel SJ, Schwan TG - PLoS Negl Trop Dis (2013)

Bottom Line: We then applied a novel genetic system for B. turicatae to inactivate brpA and examined the role of the gene product for vector colonization and the ability to establish murine infection.These results demonstrate the complexity of protein production in a population of spirochetes within the tick.Additionally, the development of a genetic system is important for future studies to evaluate the requirement of specific B. turicatae genes for vector colonization and transmission.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Mississippi State University, Starkville, Mississippi, United States of America.

ABSTRACT

Background: Borrelia turicatae, an agent of tick-borne relapsing fever, is an example of a pathogen that can adapt to disparate conditions found when colonizing the mammalian host and arthropod vector. However, little is known about the genetic factors necessary during the tick-mammalian infectious cycle, therefore we developed a genetic system to transform this species of spirochete. We also identified a plasmid gene that was up-regulated in vitro when B. turicatae was grown in conditions mimicking the tick environment. This 40 kilodalton protein was predicted to be surface localized and designated the Borrelia repeat protein A (brpA) due to the redundancy of the amino acid motif Gln-Gly-Asn-Val-Glu.

Methodology/principal findings: Quantitative reverse-transcriptase polymerase chain reaction using RNA from B. turicatae infected ticks and mice indicated differential regulation of brpA during the tick-mammalian infectious cycle. The surface localization was determined, and production of the protein within the salivary glands of the tick was demonstrated. We then applied a novel genetic system for B. turicatae to inactivate brpA and examined the role of the gene product for vector colonization and the ability to establish murine infection.

Conclusions/significance: These results demonstrate the complexity of protein production in a population of spirochetes within the tick. Additionally, the development of a genetic system is important for future studies to evaluate the requirement of specific B. turicatae genes for vector colonization and transmission.

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Related in: MedlinePlus

Amino acid alignments of BrpA from seven isolates of B. turicatae.Shaded grey are conserved amino acids, with the unshaded region depicting the Gln-Gly-Asn-Val-Glu repeats.
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pntd-0002514-g001: Amino acid alignments of BrpA from seven isolates of B. turicatae.Shaded grey are conserved amino acids, with the unshaded region depicting the Gln-Gly-Asn-Val-Glu repeats.

Mentions: Amplifying the brpA locus from the seven isolates of B. turicatae produced a product of the expected size, and the sequenced amplicons confirmed the presence of the gene in all isolates. The amino acid sequence alignment indicated high sequence identity at the amino and carboxy terminus of the protein (Figure 1). Interestingly, the alignments also revealed a redundancy in the amino acid sequence Gln-Gly-Asn-Val-Glu, with TCB-1 displaying the greatest number of repeats (Figure 1).


Development of genetic system to inactivate a Borrelia turicatae surface protein selectively produced within the salivary glands of the arthropod vector.

Lopez JE, Wilder HK, Hargrove R, Brooks CP, Peterson KE, Beare PA, Sturdevant DE, Nagarajan V, Raffel SJ, Schwan TG - PLoS Negl Trop Dis (2013)

Amino acid alignments of BrpA from seven isolates of B. turicatae.Shaded grey are conserved amino acids, with the unshaded region depicting the Gln-Gly-Asn-Val-Glu repeats.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3814808&req=5

pntd-0002514-g001: Amino acid alignments of BrpA from seven isolates of B. turicatae.Shaded grey are conserved amino acids, with the unshaded region depicting the Gln-Gly-Asn-Val-Glu repeats.
Mentions: Amplifying the brpA locus from the seven isolates of B. turicatae produced a product of the expected size, and the sequenced amplicons confirmed the presence of the gene in all isolates. The amino acid sequence alignment indicated high sequence identity at the amino and carboxy terminus of the protein (Figure 1). Interestingly, the alignments also revealed a redundancy in the amino acid sequence Gln-Gly-Asn-Val-Glu, with TCB-1 displaying the greatest number of repeats (Figure 1).

Bottom Line: We then applied a novel genetic system for B. turicatae to inactivate brpA and examined the role of the gene product for vector colonization and the ability to establish murine infection.These results demonstrate the complexity of protein production in a population of spirochetes within the tick.Additionally, the development of a genetic system is important for future studies to evaluate the requirement of specific B. turicatae genes for vector colonization and transmission.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Mississippi State University, Starkville, Mississippi, United States of America.

ABSTRACT

Background: Borrelia turicatae, an agent of tick-borne relapsing fever, is an example of a pathogen that can adapt to disparate conditions found when colonizing the mammalian host and arthropod vector. However, little is known about the genetic factors necessary during the tick-mammalian infectious cycle, therefore we developed a genetic system to transform this species of spirochete. We also identified a plasmid gene that was up-regulated in vitro when B. turicatae was grown in conditions mimicking the tick environment. This 40 kilodalton protein was predicted to be surface localized and designated the Borrelia repeat protein A (brpA) due to the redundancy of the amino acid motif Gln-Gly-Asn-Val-Glu.

Methodology/principal findings: Quantitative reverse-transcriptase polymerase chain reaction using RNA from B. turicatae infected ticks and mice indicated differential regulation of brpA during the tick-mammalian infectious cycle. The surface localization was determined, and production of the protein within the salivary glands of the tick was demonstrated. We then applied a novel genetic system for B. turicatae to inactivate brpA and examined the role of the gene product for vector colonization and the ability to establish murine infection.

Conclusions/significance: These results demonstrate the complexity of protein production in a population of spirochetes within the tick. Additionally, the development of a genetic system is important for future studies to evaluate the requirement of specific B. turicatae genes for vector colonization and transmission.

Show MeSH
Related in: MedlinePlus