title: Screening and molecular cloning of a protective antigen from the midgut of Haemaphysalis longicornis.
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 α-helical structure. These results provide the basis for developing a vaccine against biting H. longicornis ticks.
authors.affiliation: Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology of Hebei Province, College of Life Sciences, Hebei Normal University, Shijiazhuang 050024, China.
authors.forename: Yonghong
authors.initials: Y
authors.lastname: Hu
authors.forename: Jincheng
authors.initials: J
authors.lastname: Zhang
authors.forename: Shujie
authors.initials: S
authors.lastname: Yang
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Hua
authors.initials: H
authors.lastname: Zeng
authors.forename: Tiantian
authors.initials: T
authors.lastname: Zhang
authors.forename: Jingze
authors.initials: J
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D015153:Blotting, Western; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D000939:Epitopes; D005786:Gene Expression Regulation / Q000502:physiology*; D026863:Ixodidae / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D053719:Tandem Mass Spectrometry
chemical_list: D000941:Antigens; D018076:DNA, Complementary; D000939:Epitopes
keywords: Haemaphysalis longicornis; Hl-Pmy; P92; epitope; protective antigen