[Construction of cDNA expression library of salivary gland from Boophilus microplus].
Total RNA were isolated from salivary gland dissected from partially engorged Boophilus microplus. The mRNA was 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 EcoR I/Hind III arms of the lambda SCREEN vector. The recombinant phage DNA was packaged by phage-marker packaging extracts, resulting in a primary cDNA library with a size of 1.38x10(6) PFU. Data showed 100% of the library were recombinant and the titer of the amplified library was 2x10(9) PFU/ml. A partial cDNA encoding cytochrome oxidase C subunit II of B. microplus was screened from the expression library with rabbit serum against B. microplus salivary gland proteins. The results is suggested that the cDNA expression library has been constructed.
Tian
Zhan-Cheng
ZC
Key Laboratory of Veterinary Parasitology of Gansu Province, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, CAAS, Lanzhou 730046, China.
Liu
Guang-Yuan
GY
Xie
Jun-Ren
JR
Gong
Zhen-Li
ZL
D000818:Animals; D002417:Cattle / Q000469:parasitology*; D018076:DNA, Complementary / Q000235:genetics; D015723:Gene Library; D012333:RNA, Messenger / Q000235:genetics; D012469:Salivary Glands / Q000737:chemistry; D013987:Ticks / Q000235:genetics*
D018076:DNA, Complementary; D012333:RNA, Messenger