A correction in the nucleotide sequence of the Tn903 kanamycin resistance determinant in pUC4K [published erratum appears in Nucleic Acids Res 1988 Aug 11;16(15): 7762]
A correction in the nucleotide sequence of the Tn903 kanamycin resistance determinant in pUC4K [published erratum appears in Nucleic Acids Res 1988 Aug 11;16(15): 7762]
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pUC4K 中 Tn903 卡那霉素抗性决定簇的核苷酸序列的修正 [已发表的勘误表出现在 Nucleic Acids Res 1988 Aug 11;16(15): 7762]
DOI：
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发表时间：
1988
期刊：
影响因子：
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通讯作者：
R. Rose
中科院分区：
文献类型：
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作者：
L. Taylor;R. Rose
PUC4K是一种携带Tn903(1)卡那霉素抗性决定簇的载体，它不仅被用于构建新的载体，还被用作限制性内切酶动员元件，在克隆的基因(1)中产生不同长度和位置的密码子插入。PUC4K最初是通过G-C拖尾将1.4kb的Haell片段从Tn903克隆到pUC7的Pst1位点而构建的(1)。Haell片段中包含的卡那霉素抗性基因侧翼应该有226个碱基的反向重复序列(2)，但在构建PUC卡那霉素衍生物的过程中，我们发现其中一个反向重复序列缺失。通过对每个Q-C末端进行测序，我们发现该基因5*端的反向重复序列除了6个碱基外都被完全缺失。最初我们认为这个缺失是我们分离的pUC4K所独有的，但来自其他来源(Pharmacia，同事)的pUC4K包含同样的缺失。因此，pUC4K中包含的Tn903片段从碱基1052延伸到2264，而不是最初建议的碱基831到2264(2)。缺失似乎发生在pUC4K的初始构建过程中，可能是在1052碱基的缺口处拖尾。
pUC4K Is a vector that carries the kanamycin resistance determinant from Tn903 (1) and has been used not only in the construction of new vectors but also as a restriction mobilization element to create codon Insertions of varying length and location In cloned genes (1). pUC4K was originally constructed by cloning a 1.4 kb Haell fragment from Tn903 Into the Pstl site of pUC7 by G-C tailing (1). The kanamycin resistance gene contained within the Haell fragment should be flanked by 226 bp Inverted repeats (2), but in the process of constructing pUC kanamycin derivatives we have discovered that one of these Inverted repeats is missing. By sequencing through each of the Q-C tails Into the kanamycin resistance determinant we found that the inverted repeat at the 5* end of the gene had been entirely deleted except for 6 bp. Initially we thought this deletion was unique to our Isolate of pUC4K, but pUC4K from other sources (Pharmacia, colleagues) contained the same deletion. Thus, the Tn903 fragment contained in pUC4K extends from bases 1052 to 2264 instead of bases 831 to 2264 as originally suggested (2). The deletion appears to have occured during the initial construction of pUC4K, perhaps by tailing at a nick at base 1052.