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Physical mapping of a large-plaque mutation of adenovirus type 2.

机译:腺病毒2型大斑突变的物理映射。

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We have developed a simple method based on cotransfection of overlapping DNA restriction fragments for construction of recombinants of adenovirus type 2 (Ad2) and Ad5. When Ad2 DNA digested with restriction endonuclease EcoRI was cotransfected with Ad5 DNA digested with SalI, recombination occurred between Ad2 EcoRI-A (map position 0 to 59) and Ad5 SalI-A (map position 45 to 100). Analysis of the recombinant DNAs by digestion with EcoRI or BamHI restriction endonucleases indicated that, as expected, recombination had occurred in overlapping sequences (map position 45 to 59) between the Ad2 EcoRI-A fragment and the Ad5 SalI-A fragment. By using this method, several recombinants were constructed between a large-plaque (lp) mutant of Ad2 and wild-type Ad5. Cleavage of the recombinant genomes with restriction endonucleases BamHI, EcoRI, and HindIII revealed that the lp mutation is located within the left 41% of Ad2 genome.
机译:我们已经开发了一种基于重叠DNA限制片段的Cot转酵母的简单方法,用于构建腺病毒2型(AD2)和AD5的重组。当用限制性内切核酸酶消化的AD2 DNA用盐酸盐消化的AD5 DNA被分配,在AD2 EcoRI-A(MAP位置0至59)和AD5 SALI-A(MAP位置45至100)之间发生重组。通过与生态学或BamHI限制性内切核酸酶消化的消化分析,表明,如预期的那样,在AD2 EcoRI-A片段和Ad5 Sali-A片段之间的重叠序列(Map位置45至59)中发生了重组。通过使用该方法,在Ad2和野生型AD5的大斑块(LP)突变体之间构建了几种重组剂。具有限制性内切核酸酶Bamhi,EcoRI和HindIII的重组基因组的切割揭示了LP突变位于Ad2基因组的左41%内。

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