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本文报导以人活化B淋巴细胞的3D5细胞的mRNA为模板,逆转录合成cDNA的第一链:按常规的自身引物法,以合成的第一链为模板合成cDNA第、二链;以及λgt11Sfi-Not噬菌体为载体,定向插入构建了人活化B细胞。DNA表达文库。该文库的大小为2.5×106pfu,插入片段长度大于1kb;用抗CD71及CD98单克隆抗体为探针,对所构建的文库进行筛选,均获得了阳性克隆,CD71及CD98均为人B细胞活化时才得以表达的分化抗原,从而证明,新构建的cDNA文库是人活化B细胞的cDNA表达文库。
In this paper, the mRNA of human activated B lymphocytes in 3D5 cells was used as a template to reverse transcribe the first strand of cDNA: The first and second strands of cDNA were synthesized according to the conventional first-strand method as a template, and the λgt11Sfi- Not phage as a vector, directional insert constructed human activated B cells. DNA expression library. The size of the library was 2.5 × 106pfu and the length of inserted fragment was more than 1kb. Positive clones were obtained by screening the constructed libraries using anti-CD71 and CD98 monoclonal antibodies as probes. Both CD71 and CD98 were human B cells Activated when they were able to express differentiation of the antigen, thus demonstrating that the newly constructed cDNA library is a human activated B cell cDNA expression library.