【摘要】 设计“链扩增致表达框架法”改良聚合酶链扩增（PCR）技术以获得CD₄基因的V₁-V₂区段,使原不完整的真核基因区段强制性地修饰成有表达可能的完整基因,并保持原读码框架的正确性。该扩增片段经序列分析确证后,克隆入表达载体pLSD₁₃（含P_L强启动子）和pBV220（含P_RP_L融合启动子）中。Southern杂交证实了这种插入的正确性,SDS/PAGE检测诱导表达的结果,获得一个分子量为2.03×10⁴的蛋白带,证实为CD₄ V₁-V₂区蛋白。更多还原

【Abstract】 We describe the use of a modified technique of polymerase chain reaction （PCR） for facilitating cloning and expression of a cDNA fragment encoding CD₄ V₁-V₂ domains. The modification includes introducing suitable signals to start primer and halt primer of the target gene as indicated in Fig. 1. After verified by DNA sequencing, the amplified DNA fragment was cloned into prokaryotic expression vectors containing λP_L promoter. The screened clones were induced to express the target fragment V₁-V₂ of CD₄ gene and the expected gene product was estimated to be 8% or 10% of the total cellular proteins.更多还原