Wyman A R, Wolfe L B, Botstein D
Proc Natl Acad Sci U S A. 1985 May;82(9):2880-4. doi: 10.1073/pnas.82.9.2880.
The growth of clones of human genomic DNA fragments in a bacteriophage lambda vector has been examined in a number of different Escherichia coli hosts. A large proportion (8.9%) of the phages carrying different fragments of the human genome fail to grow on standard rec+ hosts but will grow on hosts carrying mutations in the recB, recC, and sbcB genes. Heteroduplex analysis in the electron microscope of DNA from four of these phages revealed substantial secondary structure, including snap-back regions 200-500 base pairs in length. Such structures were not found in phages from the same DNA library that grow in rec+ hosts. These results are interpreted in the light of prior observations [Leach, D.R.F. & Stahl, F. (1983) Nature (London) 305, 448-451] showing that inverted repetitions cloned in phage lambda can be propagated in recB recC sbcB hosts but not in rec+ hosts.
已在多种不同的大肠杆菌宿主中检测了人类基因组DNA片段克隆在λ噬菌体载体中的生长情况。携带人类基因组不同片段的噬菌体中有很大一部分(8.9%)在标准rec⁺宿主上无法生长,但能在recB、recC和sbcB基因发生突变的宿主上生长。对其中四个噬菌体的DNA进行电子显微镜异源双链分析,发现了大量二级结构,包括长度为200 - 500个碱基对的回折区域。在rec⁺宿主中生长的来自同一DNA文库的噬菌体中未发现此类结构。根据先前的观察结果[利奇,D.R.F. & 施塔尔,F.(1983年)《自然》(伦敦)305卷,448 - 451页]对这些结果进行了解释,该观察结果表明,克隆在λ噬菌体中的反向重复序列可在recB recC sbcB宿主中繁殖,但不能在rec⁺宿主中繁殖。