Korean J Parasitol.  2013 Jun;51(3):327-334. 10.3347/kjp.2013.51.3.327.

Screening and Molecular Cloning of a Protective Antigen from the Midgut of Haemaphysalis longicornis

Affiliations
  • 1Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology of Hebei Province, College of Life Sciences, Hebei Normal University, Shijiazhuang 050024, China. jzliu21@heinfo.net
  • 2Shijiazhuang Post and Telecommunication Technical College, Shijiazhuang 050021, China.

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.

Keyword

Haemaphysalis longicornis; protective antigen; P92; Hl-Pmy; epitope

MeSH Terms

Amino Acid Sequence
Animals
Antigens/genetics/*metabolism
Base Sequence
Blotting, Western
Chromatography, Liquid
*Cloning, Molecular
DNA, Complementary/genetics
Epitopes
Gene Expression Regulation/*physiology
Ixodidae/genetics/*metabolism
Molecular Sequence Data
Tandem Mass Spectrometry
Antigens
DNA, Complementary
Epitopes
Full Text Links
  • KJP
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
    DB Error: unknown error