B细胞活化基因的研究Ⅰ．人活化B细胞cDNA文库的构建

Studies on B Cell Activation Genes I.Construction of Human Activated B Cell cDNA Library

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。

n order to carry out the studies on the structures and functions of B cell acti- vation genes and their products, we used the mRNA of a human activated B cellline, 3D5,as template to synthesize its cDNA through reverse- transcription, ligatedthe cDNA directionally with λgt11 Sfi-Not vector arms,and thus constructed ahuman activated B lymphocyte cDNA expressive library. The size of our cDNA library is 2.5 ×106pfu,The length of the cDNA insertsis longer than 1kb.Possitive clones are obtained when the library is screened withanti-CD71 and anti-CD98 monoclonal antibodies.As it is well known that theCD71 and CD98 are both differentiation antigens which appear onto the cellsurface only after the cell has been activated,we conclude that the cDNA librarywe have constructed is a human activated B lymphocyte cDNA expressive library.