Zaballos A, Salas M, Mellado R P
Centro de Biologia Molecular (CSIC-UAM), Universidad Autónoma, Canto Blanco, Madrid, Spain.
Gene. 1987;58(1):67-76. doi: 10.1016/0378-1119(87)90030-8.
A set of plasmid expression vectors for cloning of DNA fragments containing open reading frames has been obtained. The plasmids carry the strong leftward promoter of bacteriophage lambda and the translation initiation signals from either the gene ner of bacteriophage Mu or the gene 4 of bacteriophage phi 29. The vectors could overexpress the cloned sequences as fusion peptides at the N terminus with the N-terminal segment of the phi 29 protein p4 or at the C terminus with the Escherichia coli beta-galactosidase from its 8th residue, or both. Alternatively, the cloned sequences could be directed to overproduce proteins in an unfused form. DNA fragments of the hemagglutinin gene from human influenza A virus, have been cloned in one of the plasmid vectors and some potential antigenic determinants have been characterized using monoclonal antibodies.
已获得一组用于克隆含有开放阅读框的DNA片段的质粒表达载体。这些质粒携带噬菌体λ的强向左启动子以及来自噬菌体Mu的ner基因或噬菌体φ29的基因4的翻译起始信号。这些载体可以将克隆的序列作为融合肽在N端与φ29蛋白p4的N端片段过表达,或在C端与大肠杆菌β-半乳糖苷酶从其第8个残基起过表达,或两者同时过表达。或者,克隆的序列可以被定向以未融合的形式过量产生蛋白质。来自人类甲型流感病毒血凝素基因的DNA片段已被克隆到其中一种质粒载体中,并且使用单克隆抗体对一些潜在的抗原决定簇进行了表征。