The Construction And Immuno-Screening Of CDNA Expression Library Of Haemaphysalis Qinghaiensis Larval Tick

Heamaphysalis qinghaiensis is an ectoparasite firstly named in China. Ittransmits many species of haemoprotozoa parasitese except that it decreases theproducity of the animals by biting them to suck their blood. It inhibits the development of animal husbandry in China.By this essay, a cDNA Expression Library from Haemaphysalis qinghaiensis larval ticks was constructe. The primary library capacitance is 5.0×10~5 PFU/ml and the amplification library capacitance is 8×10~9 PFU/ml. The library was screened with positive serum of rabbit against Haemaphysalis qinghaiensis larval tick, the positive plaques were sub-cloned, their plasmids were extracted and transformed to JM109. 11 genes were obtained, and they were named as HqL01, HqL02, HqL03, HqL04, HqL05, HqL06, HqL07, HqL08, HqL09, HqL10, HqL11. The result of comparation to known gene sequences revealed that six of them were new genes. The sequence dates of HqL07, HqL08, HqL09, HqL11 were submited to GenBank, accession numbers provided by them were EF605263, EF605264, EF605265, EF605266. Three of them were expressed, they were gene HqL07, HqL08, HqL09 and maybe have good immunogenicity. Molecular weight of expression products were 60kD, 35kD, 70kD. Analysis of western blot essay show expression product of gene HqL07, which molecular weight is 60kD, has very good immunogenicity.