Ferrari E, Henner D J, Hoch J A
J Bacteriol. 1981 Apr;146(1):430-2. doi: 10.1128/jb.146.1.430-432.1981.
A library of Bacillus subtilis chromosomal deoxyribonucleic acid (DNA) was constructed, using lambda charon 4A as a cloning vector. Partially cleaved Bacillus subtilis DNA was prepared by partial methylation with EcoRI methylase, followed by complete EcoRI endonuclease digestion. More than 95% of the phage particles carried B. subtilis DNA inserts. When this library was screened for transforming activity, using competent cells, 70% of the genetic markers tested were found in a sample of 1,710 plaques. Cloned genetic loci were found to be about 100-fold more efficient in transforming activity than chromosomal DNA. Intact phage particles containing the pheA locus were found to be able to transform competent recipients with approximately the same efficiency as phage DNA. Transformation by intact particles was insensitive to deoxyribonuclease.
构建了枯草芽孢杆菌染色体脱氧核糖核酸（DNA）文库，使用λ噬菌体载体Charon 4A。通过用EcoRI甲基化酶进行部分甲基化，随后进行完全的EcoRI核酸内切酶消化来制备部分切割的枯草芽孢杆菌DNA。超过95%的噬菌体颗粒携带枯草芽孢杆菌DNA插入片段。当使用感受态细胞对该文库进行转化活性筛选时，在1710个噬菌斑样本中发现70%的测试遗传标记。发现克隆的基因座在转化活性方面比染色体DNA高效约100倍。发现含有pheA基因座的完整噬菌体颗粒能够以与噬菌体DNA大致相同的效率转化感受态受体。完整颗粒介导的转化对脱氧核糖核酸酶不敏感。
