Preliminary Analysis The Immune Protective Of Haemaphysalis Longicornis Tm Protein And MALC Protein

Ticks transmitted diseases pathogenic ability, the world’s second, is one of the most important zoonosis media, especially the Forest Encephalitis, Xinjiang Hemorrhagic Fever, Lyme disease and Q fever caused by pathogens transmitted by the hard ticks, which seriously threat the health of mankind and the development of husbandry. The Safe, no pollution, effective prevention and control of ticks is emergent question needed to be solved. During the recent years, the use of vaccines to biological tick control become one of the most promising prevention methods ticks, while this approach mainly depends on the identification of protective antigens in ticks.Bioinformatics analysis the positive clones which was screened from a cDNA library constructed from partial-fed female ticks, and then cloned and expressed HL-Tm gene and HL-MALC gene. The expression and reactogenicity of recombinant protein were analyzed by SDS-PAGE and Western-blot. Real-time comparative quantitative RT-PCR analysis of HL-MALC and HL-Tm in various tissues and in different developmental stages. Immunization and challenge infestation and RNAi of unfed and starved ticks to assess its feasibility for tick control immune vaccine candidate antigen. To offer the platform for investigating the molecular mechanism of controlling on the physiological behavior of ticks and evaluating the neotype environmentally friendly pest biological agents and furthermore make up the deficiency in the intervention of physiological behavior and the controlling neotype agent. The mostly results as followed.The HL-MALC gene and HL-Tm gene was cloned into pGEX-4T-1vector and was expressed in Escherichia coli BL21(DE3) respectively. Western blotting showed that rabbit anti-serum against H. longicornis unfed-adult ticks could produce the significant band with rHL-MALC-GST protein.Real-time RT-PCR analysis showed that the HL-MALC transcripts were expressed in different developmental stages of H. longicornis ticks and were significantly expressed at the unfed-female ticks. Furthermore, the expression of HL-MALC and HL-Tm in epidermis was significantly higher than that of other tested tissues of partially fed female ticks, because ticks muscles mainly adhesion in the skin inside.Vaccination of rabbits with the rHL-MALC and rHL-Tm conferred partial immunological protective effects against adult ticks. The results of RNAi experiment were also confirmed that the HL-MALC gene and HL-Tm gene played an important roles in the regulation of physiological process of H. longicornis.