莫氏巴贝斯虫裂殖子cDNA表达文库的构建及免疫学筛选

Abstract

Abstract:

The objective of this study was to obtain functional genes of Babesia motasi, a cDNA expression library of the merozoites was constructed and immunoscreened with positive sera from sheep infected with B. motasi. The merozoites of B. motasi were purified from red blood cell with differential centrifugation. The mRNA was purified from extracted total RNA. Synthetized double-strand cDNA was added directional EcoRⅠ/Hind Ⅲ linkers and ligated to the EcoRⅠ/HindⅢ arms of λ screen vector. To produce a primary cDNA library of B. motasi, the phages DNA was packaged in vitro and transfected into ER1647. The positive clones were obtained by immunoscreening with the positive sera against B. motasi and amplified with rapid amplification of cDNA ends (RACE). The titers of the primary and amplified cDNA expression library were 1.0×10⁶PFU and 3.5×10⁹ PFU·mL^-1, respectively. The results showed that 10 genes of B. motasi were identified and the full-length of 8 genes were amplified by RACE. The cDNA expression library and genes screened provide an important material for screening and identifying candidate antigens of vaccination and diagnosis, drug targets as well studying biological characteristics of Babesia.

CLC Number:

S852.723

WANG Jin-ming, LIU Jun-long, LIU Ai-hong, MA Mi-ling, NIU Qing-li, REN Qiao-yun, YANG Ji-fei, LIU Zhi-jie, LI You-quan, LUO Jian-xun, YIN Hong, GUAN Gui-quan. Construction and Immunoscreening of a Merozoites cDNA Expression Library of Babesia motasi[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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Construction and Immunoscreening of a Merozoites cDNA Expression Library of Babesia motasi

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