In order to understand the structure and function of fatty acid binding proteins(FABP)and the genes in ticks,primers were designed based on the sequence of Contig879 in the transcriptome library of Haemaphysalis flava,and the missing sequences of the two ends were amplified by the rapid-amplification of cDNA ends(RACE)method.Real-time PCR(qPCR)was used to determine the transcription level of the gene in different developmental stages and organs of ticks.The effect of H.flava FABP(HfFABP)dsRNA interference on the protoplasmic proteins of the eggs was analyzed by the parallel reaction monitoring(PRM)method.The results showed that the full-length HfFABP cDNA was 1443bp,and the ORF was 396bp,and it encoded a polypeptide of 132aa.The transcription level of this gene in female ticks was higher than that in larvae,nymphs and male ticks;the transcription level in the ovary,midgut,and integument was higher than that in the salivary gland.Blood sucking increased the transcription level of HfFABP.The interference of HfFABP dsRNA reduced spawning,and about 30%of the eggs were discrepant in size,waxy,fragile and easy to crack.The hatching rate of apparently normal eggs was reduced.The abundance of many proteins in eggs,such as FABP,vitellogenin,neutrophil elastase inhibitor,aspartic protease,and heat shock protein 83,decreased significantly in ticks with HfFABP dsRNA interference(P<0.01,P<0.05).These results indicate that HfFABP plays an important role in the development of tick eggs.This study provides a scientific basis for the application of HfFABP in the anti-female tick reproduction.
Haemaphysalis flavafatty acid-binding proteins(FABP)rapid-amplification of cDNA ends(RACE)parallel reaction monitoring(PRM)
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