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Published on: July 23, 2017
Characterization of a Vitellogenin Gene Fragment in Boophilus microplus Ticks
Gabriela Granjeno-Colin1, Ruben Hernandez-Ortiz, Juan Mosqueda
1FMVZ, Universidad Nacional Autonoma de México, D.F. México, México.
Abstract:
The objective of this study was to isolate, clone, and characterize a fragment of the vitellogenin (Vg) gene from a B. microplus tick strain from Mexico. Using cDNA and specific primers, an 1800-bp fragment was amplified, cloned, and transformed in into E. coli, and then sequenced. Comparative analysis with a previously reported sequence showed 99% identity at both the nucleotide and amino acid level. The predicted amino acid sequence of the Mexican Vg has 6 positive mutations. There is an insertion of an aspartic acid on position 26 and a deletion on position 552 with respect to the reported sequence. There were 11 predicted glycosylation sites conserved in both strains. It is concluded that there is a high sequence homology of Vg in both strains.
Insights
Researchers isolated and sequenced a vitellogenin (Vg) gene fragment from Mexican B. microplus ticks. The gene shows high similarity to other strains, indicating conserved function in this important cattle parasite.
Area of Science:
- Molecular Biology
- Parasitology
- Genomics
Background:
- Vitellogenin (Vg) is crucial for egg development in arthropods.
- Understanding Vg gene variations in ticks like *Boophilus microplus* (B. microplus) is key to developing control strategies.
- Genetic diversity in Vg can impact reproductive fitness and insecticide resistance.
Purpose of the Study:
- To isolate, clone, and characterize a vitellogenin (Vg) gene fragment from a Mexican *B. microplus* tick population.
- To compare the genetic sequence and protein structure with previously reported Vg sequences.
- To identify potential genetic variations and conserved functional sites.
Main Methods:
- RNA extraction from *B. microplus* ticks.
- Complementary DNA (cDNA) synthesis.
- Polymerase Chain Reaction (PCR) amplification using specific primers.
- Gene cloning into *Escherichia coli* (E. coli).
- DNA sequencing and comparative bioinformatic analysis.
Main Results:
- An 1800-bp fragment of the *B. microplus* Vg gene was successfully amplified and sequenced.
- The Mexican Vg sequence exhibited 99% identity at both nucleotide and amino acid levels compared to a reference sequence.
- Six positive mutations, including an aspartic acid insertion and a deletion, were identified in the Mexican Vg sequence.
- Eleven predicted glycosylation sites were conserved between the Mexican and reference Vg strains.
Conclusions:
- The *B. microplus* Vg gene in the Mexican strain shows high sequence homology with other reported strains.
- Conserved glycosylation sites suggest functional importance and potential conservation of Vg protein function.
- Identified mutations may offer insights into potential adaptations or variations within *B. microplus* populations.
