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Screening and Molecular Cloning of a Protective Antigen from the Midgut of Haemaphysalis longicornis
Article in English | WPRIM | ID: wpr-79739
Responsible library: WPRO
ABSTRACT
Vaccination is considered a promising alternative for controlling tick infestations. Haemaphysalis longicornis midgut proteins separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membrane were screened for protective value against bites. The western blot demonstrated the immunogenicity of 92 kDa protein (P92). The analysis of the P92 amino acid sequence by LC-MS/MS indicated that it was a H. longicornis paramyosin (Hl-Pmy). The full lenghth cDNA of Hl-Pmy was obtained by rapid amplification of cDNA ends (RACE) which consisted of 2,783 bp with a 161 bp 3' untranslated region. Sequence alignment of tick paramyosin (Pmy) showed that Hl-Pmy shared a high level of conservation among ticks. Comparison with the protective epitope sequence of other invertebrate Pmy, it was calculated that the protective epitope of Hl-Pmy was a peptide (LEEAEGSSETVVEMNKKRDTE) named LEE, which was close to the N-terminal of Hl-Pmy protein. The secondary structure analysis suggested that LEE had non-helical segments within an alpha-helical structure. These results provide the basis for developing a vaccine against biting H. longicornis ticks.
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Full text: Available Index: WPRIM (Western Pacific) Main subject: Molecular Sequence Data / Base Sequence / Gene Expression Regulation / Blotting, Western / Chromatography, Liquid / Amino Acid Sequence / Cloning, Molecular / DNA, Complementary / Ixodidae / Tandem Mass Spectrometry Type of study: Screening study Language: English Journal: The Korean Journal of Parasitology Year: 2013 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Molecular Sequence Data / Base Sequence / Gene Expression Regulation / Blotting, Western / Chromatography, Liquid / Amino Acid Sequence / Cloning, Molecular / DNA, Complementary / Ixodidae / Tandem Mass Spectrometry Type of study: Screening study Language: English Journal: The Korean Journal of Parasitology Year: 2013 Type: Article