本文报道以ＴＰＡ活化Ｊｕｒｋａｔ细胞ｍＲＮＡ为模板，逆转录合成ｃＤＮＡ第１链，在ＤＮＡ聚合酶Ⅰ作用下合成第２链，加上ＥｃｏＲＩ磷酸化接头，用离心式柱层析去除小于５００ｂｐ的ｃＤＮＡ片段和多余接头，再与去磷酸化λｇｔ（１１）ＤＮＡ连接，体外包装后感染大肠杆菌Ｙ１０９０。该文库效价５×１０５，重组率＞９０％。ＰＣＲ法鉴定阳性重组子均可见插入片段。用ＰＴＡ１ＭｃＡｂ为探针，对文库进行了免疫学筛选，初步获得４个阳性克隆。该文库的构建为ＰＴＡ１基因克隆奠定了基础。 In this paper, TPA-activated Jurkat cell mRNA was used as a template to reverse transcribe the first strand of cDNA. The second strand was synthesized under the action of DNA polymerase I and EcoRI phosphorylated linker was used to remove cDNA fragments smaller than 500 bp by centrifugal column chromatography And the extra linker, and then connected with the dephosphorylated λgt (11) DNA, packaged in vitro infection Escherichia coli Y1090. The library titer 5 × 105, recombination rate> 90%. Positive recombinant PCR were identified insert can be seen. Using PTA1McAb as probe, the library was screened by immunology and four positive clones were obtained. The construction of this library laid the foundation for the cloning of PTA1 gene.