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两种新工具:携带卡那霉素或壮观霉素/链霉素抗性标记的多用途克隆载体。

Two new tools: multi-purpose cloning vectors that carry kanamycin or spectinomycin/streptomycin resistance markers.

作者信息

Kirschman J A, Cramer J H

机构信息

Agrigenetics Advanced Science Company, Madison, WI 53716.

出版信息

Gene. 1988 Aug 15;68(1):163-5. doi: 10.1016/0378-1119(88)90609-9.

Abstract

We have constructed two new multi-purpose cloning vectors, pJKKmf(-) and pJKSp/Smf(-), that carry resistance to kanamycin (Km) and spectinomycin/streptomycin (Sp/Sm), respectively. These plasmids are based on pGEM3Zf(-) and therefore contain a pUC-vector-derived multiple cloning site for 13 restriction sites flanked by T7 and SP6 RNA polymerase promoters, the alpha-peptide coding region of beta-galactosidase for blue/white colony screening and the bacteriophage f1 origin of replication for production of single-stranded DNA in the presence of a helper phage. We have used these vectors to reclone sequences from a maize genomic library, to produce radiolabeled RNA probes and to make single-stranded DNA.

摘要

我们构建了两种新型多用途克隆载体,即pJKKmf(-)和pJKSp/Smf(-),它们分别携带对卡那霉素(Km)和壮观霉素/链霉素(Sp/Sm)的抗性。这些质粒基于pGEM3Zf(-),因此包含一个源自pUC载体的多克隆位点,该位点有13个限制酶切位点,两侧为T7和SP6 RNA聚合酶启动子、用于蓝/白菌落筛选的β-半乳糖苷酶α-肽编码区以及用于在辅助噬菌体存在下生产单链DNA的噬菌体f1复制起点。我们已使用这些载体从玉米基因组文库中重新克隆序列、生产放射性标记的RNA探针并制备单链DNA。

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