Construction of cDNA Expression Library of Haemaphysalis qinghaiensis

Total RNA were isolated from organs such as salivary glands,Malpighian tubules, ovaries dissected from partially engorged Haemaphysalis qinghaiensis.Subsequently mRNA were purified.A library of oligo(dT)-primed cDNA with added directional EcoR I/Hind III linkers was constructed from the purified mRNA.The constructed cDNA was ligated to the EcoRI/HindIII arms of the λSCREEN vector.The recombinant phage DNA was packaged by using PhageMaker packaging extracts,resulting in a primary cDNA library with a size of 2.0×10~6 PFU.Data showed 100% of the library were recombinant and the titer of the amplified library was 8×10~9PFU.A full-length cDNA encoding myosin light chain alkali of Haemaphysalis qinghaiensis was screened from the expression library using rabbit anti-Haemaphysalis qinghaiensis differential proteins serum.The results suggested that the cDNA expression library was constructed successfully.