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Characterization and molecular cloning of cryptic plasmids isolated from Lactobacillus casei.

机译:从干酪乳杆菌分离的隐性质粒的表征和分子克隆。

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摘要

Four small cryptic plasmids were isolated from Lactobacillus casei strains, and restriction endonuclease maps of these plasmids were constructed. Three of the small plasmids (pLZ18C, pLZ19E, and pLZ19F1; 6.4, 4.9, and 4.8 kilobase pairs, respectively) were cloned into Escherichia coli K-12 by using pBR322, pACYC184, and pUC8 as vectors. Two of the plasmids, pLZ18C and pLZ19E, were also cloned into Streptococcus sanguis by using pVA1 as the vector. Hybridization by using nick-translated cloned 32P-labeled L. casei plasmid DNA as the probe revealed that none of the cryptic plasmids had appreciable DNA-DNA homology with the large lactose plasmids found in the L. casei strains, with chromosomal DNAs isolated from these strains. Partial homology was detected among several plasmids isolated from different strains, but not among cryptic plasmids isolated from the same strain.
机译:从干酪乳杆菌菌株中分离出四个小的隐性质粒,并构建了这些质粒的限制性核酸内切酶图。通过使用pBR322,pACYC184和pUC8作为载体,将三个小质粒(分别为pLZ18C,pLZ19E和pLZ19F1;分别为6.4、4.9和4.8千碱基对)克隆到大肠杆菌K-12中。通过使用pVA1作为载体,还将两个质粒pLZ18C和pLZ19E克隆到血链球菌中。通过使用缺口翻译的克隆的32P标记的干酪乳杆菌质粒DNA进行杂交,发现没有一个隐性质粒与干酪乳杆菌菌株中发现的大乳糖质粒具有明显的DNA-DNA同源性,并从这些质粒中分离出了染色体DNA。株。在从不同菌株分离的几种质粒中检测到部分同源性,但在从同一菌株分离的隐性质粒中未检测到。

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