Identification Of Two Peptides From The Salivary Glands Of Hyalomma Asiaticum

Tick salivary has great effect on the process of feeding.it can help the tick engorge,suppress the inflammation and transmit various of diseases. Base on the cDNA library we have built, differently expressed molecular P18and Ha-11was picked up.This study has cloned expressed and the studied the primary function of Ha-11.It has an372bp ORF,encode a peptide of123aa with a single peptide of23aa. With IPTG induction,the37kDa recombinant protein was expressed by sub-cloning to the pGEX-4T-1E.coli expression vector.After injecting the mouse with recombinant protein with Freund adjuvant, antisera was made from the mouse.the anti-sera recognise a11kDa native protein in the salivary of tick.Real-time PCR showed that the molecular Ha-11was abundant in the nymphae and Hemolymph.recombinant protein can suppress the proliferation of spleen cell of mouse and the expression of IL-2, IFN-γ in the supernatant of spleen cell cultural.RNAi indicated that Ha-11has limited affection on the biological characteristics.For the molecular P18, it has.an ORF of565bp,encoded155aa,has a singal of22aa.the P18was subcloned to the pET-32a E.coli expression vector.the anti-sera recognized a18kDa native protein in tick salivary.real-time PCR showed P18was mostly expressed in the nymphae and salivary protein. Recombinant protein suppressed the proliferation of spleen cell and the expression of TNF-α in the supernatant of spleen cell cultural.Knocking down the molecular in the nymphae,it has effect on the engorgement and repletion rates in ticks.The experiment has proved that the two molecular has great effects on the process of engorgement.