摘要
采用聚合酶链反应(PCR)技术，直接从微生物基因组中扩增卡介苗Ag85-B和HBsAg编码基因。虽然在引物5'端设计了限制性内切酶识别顺序以及附加顺序，但因限制性内切酶不能切开这些顺序，使扩增基因产物不能克隆到质粒载体中。因此，我们构建了新的克隆载体即T载体，专门用于克隆未经任何修饰的PCR产物；这种方法价廉，快速有效，值得推广应用。
Genes encoding BCG Ag85-B and HBsAg were amplificated directly from mi-crobiotic genome by using PCR.Although restrication endonuclease sites are often incorpo-rated into the oligonucleotide primers used for amplication,so that cleavage of the product will creat sticky ends that can theoretically be ligated to an equivalently cut vector.Unfortu-nately,this cloning way could not always lead to the successful results.Therefore,a new cloning vector(T-vector)was constructed,and specifically used to clone unmodified PCR products.This method is very economical,efficient and helpful in the study of genetic engi-neering.
出处
《同济医科大学学报》
CAS
CSCD
北大核心
1994年第3期167-170,共4页
Acta Universitatis Medicinae Tongji
基金
国家自然科学基金
关键词
聚合酶链反应
克隆
T载体
基因
polymerase chain reaction,cloning,molecular,T-vector
