中文摘要：

目的：构建β-地中海贫血CD41—42突变基因和基因调控区HS2、HS3全基因片段的真核表达重组载体，为B一地中海贫血CD41—42突变基因治疗的细胞模型奠定基础。方法：设计合成β-地中海贫血CD41—42突变纯合子患者的DNA特异性引物，扩增基因座控制区（LCR）和CD41—42突变基因，插入载体pcDNA3．1-中，筛选阳性重组体，通过酶切分析及PCR进行鉴定。结果：扩增出2．1kbCIMl—42突变基因及5．5kbLCR片段，酶切及测序结果证明重组载体构建成功，测序结果和引入方向正确。结论：成功构建了含人B一地中海贫血CD41—42基因的重组载体。

英文摘要：

Objective： To construct recombinant vector pcDNA3 functioning as a eukaryotic expression vector and carrying the CD41 -42 mutation responsible for human ~ -thalassemia. The locus control region （LCR）, which consisting of HS2, HS3, and the CD4I -42 mutation -containing fragment were used as the target fragments, respectively. This work provides the foundation for the building of ccll model and a transgenic mouse model for the gene therapy of human [3 -thalassemia. Methods： DNA extraction was performed in eight 13 -thalassemia patients who were confirmed to carry the homozygons CD4I -42 mutation. The specific primers were designed for PCR am- plification of 13- globin gene LCR and CD41 -4 mutation- containing fragment. The amplified products were directionally inserted into the eukaryotic expression vector pcDNA3.1. The positive recombinant vector was screened by restriction enzyme analysis and PCR identification. Results： Specific LCR and CD41 -42 mutation - containing fragment were amplified, with the length of approximately 5.5 kb and 2. 1 kb respectively. Restriction enzyme analysis and sequencing results indicated that the recombinant vector contained the 13 - globin gene LCR and CD41 -42 mutation -containing fragment, which were cloned successfully at the desired position. Conclusion： The recombi- nant vector carrying human [3 -globin gene LCR and CD41 -42 mutation- containing fragment is successfully constructed.