关键词: 长角血蜱, 卵, cDNA表达文库, 构建

Abstract: Abstract: 【OBJECTIVE】The cDNA Expression Library, based on the source of eggs of Haemaphysalis longicornis, was constructed successfully for further screening the potential candidate antigens angist the Haemaphysalis longicornis. 【METHOD】Total RNA were isolated from eggs of Haemaphysalis Longicornis, subsequently mRNA were purified a library of oligo(dT)-primed cDNA with added directional EcoRⅠ/Hind Ⅲ linkers was constructed from the purified mRNA. The constructed cDNA was ligated to the EcoRⅠ/Eind Ⅲ arms of the λSCREEN vector. 【RESULT】The recombinant phage DNA was packaged by using Phagemarker packaging extracts, resulting in a primary cDNA library with a size of 1.38×106 PFU. Data showed 100% of the library were recombinant and the titer of the amplified library was 2×109 PFU. A cDNA encoding follistatin-related protein was cloned from the expression library. 【CONCLUSION】The results suggested that the cDNA expression library was constructed successfully.

Key words: Haemaphysalis Longicornis, eggs, cDNA expression library, construction