Del Sal G, Manfioletti G, Gustincich S, Ruaro E, Schneider C
Laboratorio Nazionale Consorzio Interuniversitario per le Biotecnologie, Trieste, Italy.
Biotechniques. 1994 Jan;16(1):134-8.
This report describes the construction of a new family of lambda phage and plasmid cloning vectors. lambda GDST3/T7 allows cDNA insertion up to 14 kb; it is derived from lambda NM1151 by the insertion of a multiple cloning site containing eight unique restriction sites. The two asymmetrical SfiI sites are flanked by the T3 and T7 promoters for direct sequencing and in vitro transcription/translation. The same multiple cloning site is also present in both orientations in the eukaryotic expression plasmids, pGDSV3 and pGDSV7. By exploiting the superior discrimination of the signal-to-noise ratio of the lambda vectors for primary screening (by either nucleic acids or antibody probes), relevant cDNAs can thus be efficiently transferred through SfiI sites into the plasmids pGDSV3/7 for functional secondary screening by expression in mammalian cells.
本报告描述了一个新的λ噬菌体和质粒克隆载体家族的构建。λGDST3/T7允许插入长达14 kb的cDNA；它是通过插入一个含有八个独特限制酶切位点的多克隆位点从λNM1151衍生而来。两个不对称的SfiI位点两侧是T3和T7启动子，用于直接测序和体外转录/翻译。真核表达质粒pGDSV3和pGDSV7中也以两种方向存在相同的多克隆位点。通过利用λ载体在初次筛选（通过核酸或抗体探针）中卓越的信噪比辨别能力，相关的cDNA因此可以通过SfiI位点有效地转移到质粒pGDSV3/7中，以便通过在哺乳动物细胞中的表达进行功能性二次筛选。
