Studies On Gene Clone And Expression Of Haemaphysalis Flava HSP70 And Identification Of The Microflora In The Midgut

In order to provide a new view to prevent and treat the parasitic disease,the study takes Haemaphysalis flava as experimental material and researches the microflora in the midgut content,gene full-length and expression of HSP70. In the experiment,the females and males in different development stages from three areas are identified to follow-up experiments.1?We obtain the full-length cDNA sequence of HSP70 gene with RACE and analyze the various bioinformations of it. The result shows that the full-length of HSP70 cDNA sequence is 2349bp to encode a protein of 648 amino acids with a calculated molecular weight of 71164.46Da and a theoretical isoelectric point of 5.43. The HSP70 in H. flava has the highest homologous identity with Amblyomma variegatum.2?The mRNA relative expression of HSP70 gene in H. flava is analyzed by RT-PCR that take the ?-actin as reference gene. The consequences are that expression levels of HSP70 in salivary gland is higher than mid-gut and expression levels of HSP70 in partially fed female tested tissues is higher than feding males. In addition,expression of HSP70 gradually increased to achieve the maximum(engorgement females) with blood-feeding. It is suggested that HSP70 is involved with blood-feeding,digestion,immunity to the pathogeny besides cold or heat stress.3?PCR-DGGE analysis showed that 13 kinds of microbes are identified in the the midgut content from H.flava which are Rickettsia peacockii, Candidatus Rickettsia angustus, Candidatus Rickettsia kingi, Enterobacter, Coxiella burnetii, Pseudomonas aeruginosa, Klebsiella pneumoniae, Bacillus cereus, Erwinia chrysanthemi, Staphylococcus simulans, Pectobacterium carotovorum, etc. This study is significant for tick-borne diseases researches and public health.