羊尤氏泰勒虫一个新生多肽复合物α多肽基因的cDNA文库筛选和测序
양우씨태륵충일개신생다태복합물α다태기인적cDNA문고사선화측서
MONOCLONAL ANTIBODY SCREENING AND SEQUENCING OF A NASCENT POLYPEPTIDE ASSOCIATED COMPLEX SUBUNIT ALPHA-LIKE SEQUENCE FROM THEILERIA UILENBERGI CDNA LIBRARY
  • 万方数据
    中国动物传染病学报 중국동물전염병학보
    CHINESE JOURNAL OF VETERINARY PARASITOLOGY
    2010年 4期 60-65 ,共6页

    党志胜%刘志杰%罗建勋%关贵全%马米玲%刘爱红%李有全%高金亮%任巧云%白启%杉本千寻%殷宏

    당지성%류지걸%라건훈%관귀전%마미령%류애홍%리유전%고금량%임교운%백계%삼본천심%은굉

    尤氏泰勒虫%裂殖子%cDNA文库%单克隆抗体%测序%新生多肽相关复合物 尤氏泰勒蟲%裂殖子%cDNA文庫%單剋隆抗體%測序%新生多肽相關複閤物
    우씨태륵충%렬식자%cDNA문고%단극륭항체%측서%신생다태상관복합물
    Theileria uilenbergi%merozoite%cDNA library%mAb%sequencing%nascent polypeptide associated comple (NAC)
    为筛选羊尤氏泰勒虫裂殖子功能基因,用羊尤氏泰勒虫裂殖子提取物免疫BALB/c小鼠,将小鼠脾脏细胞与鼠骨髓瘤SP2/0-Ag14细胞融合,融合细胞培养上清用酶联免疫吸附法和免疫印记法检测.提取羊尤氏泰勒虫裂殖子mRNA 后,进行cDNA 合成与文库构建.文库在免疫学噬斑筛选后,将阳性克隆进行测序和序列BLAST搜索.筛选结果显示有两个阳性克隆.所得序列BLAST搜索结果显示,它与12类寄生虫新生多态相关复合物α多肽基因的同源性在39%到78%之间.结果提示,所筛选的序列为羊尤氏泰勒虫裂殖子新生多态相关复合物α多肽基因不完全序列.该研究为今后使用单克隆抗体筛选羊泰勒虫功能性基因提供了参考.
    위사선양우씨태륵충렬식자공능기인,용양우씨태륵충렬식자제취물면역BALB/c소서,장소서비장세포여서골수류SP2/0-Ag14세포융합,융합세포배양상청용매련면역흡부법화면역인기법검측.제취양우씨태륵충렬식자mRNA 후,진행cDNA 합성여문고구건.문고재면역학서반사선후,장양성극륭진행측서화서렬BLAST수색.사선결과현시유량개양성극륭.소득서렬BLAST수색결과현시,타여12류기생충신생다태상관복합물α다태기인적동원성재39%도78%지간.결과제시,소사선적서렬위양우씨태륵충렬식자신생다태상관복합물α다태기인불완전서렬.해연구위금후사용단극륭항체사선양태륵충공능성기인제공료삼고.
    Extracts of Theileria uilenbergi merozoites were used to immunize Balb/c mice and the spleen cells were fused with mouse myeloma SP2/0-Ag14. The culture supernatants were tested for antibody production by ELISA and Western blot. The monoclonal antibodies were used to screened cDNA library of T.uilenbergi merozoites. Plaque screening was carried out with the pico BlueTM Immunoscreening Kit. The positive clones were sequenced and a BLAST search was performed using the obtained sequences. Two positive clones were identified. The BLAST search revealed 39% to 78% homology of the cloned sequences with 12 parasite NAC subunit alpha sequences. This suggested that the sequence screened by mAb was the NAC subunit alpha-like partial sequence of T.uilenbergi. This is the first report on the screening of T.uilenbergi functional antigen gene by using monoclonal antibodies.
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