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CD_4基因的V_1-V_2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pLSD_(13)(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2.03×10~4的蛋白带,证实为CD_4 V_1-V_2区蛋白。
We describe the use of a modified technique of polymerase chain reaction (PCR) for facilitating cloning and expression of a cDNA fragment encoding CD_4 V_1-V_2 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 V_1-V_2 of CD_4 gene and the expected gene product was estimated to be 8% or 10% of the total cellular proteins.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
1992年第2期79-84,共6页
Acta Academiae Medicinae Sinicae
