Rossi M, Brigidi P, Matteuzzi D
Department of Pharmaceutical Sciences, University of Bologna, Italy.
Lett Appl Microbiol. 1998 Feb;26(2):101-4. doi: 10.1046/j.1472-765x.1998.00285.x.
The recombinant plasmids pDLI41, pDGA7 and pDCO7 were constructed by cloning in pDG7, a vector based on Bifidobacterium longum replicon pMB1, the following heterologous genes: Pseudomonas fluorescens lipase, Bacillus licheniformis alpha-amylase and Streptomyces sp. cholesterol oxidase. The hybrid plasmids efficiently transformed Bifidobacterium belonging to five different species. A novel Escherichia coli-Bifidobacterium set of shuttle vectors based on the replicon pMB1 (pLF5, pCLJ15, pSPEC1) featuring chloramphenicol, erythromycin and spectinomycin resistance genetic determinants as selection marker for bifidobacteria, was developed. The plasmid pTRE3, a derivative of pLF5, was the smallest (2.8 kb) Bifidobacterium vector, possessed a convenient multicloning site and presented high structural and segregational stability.
通过将以下异源基因克隆到基于长双歧杆菌复制子pMB1的载体pDG7中，构建了重组质粒pDLI41、pDGA7和pDCO7：荧光假单胞菌脂肪酶、地衣芽孢杆菌α淀粉酶和链霉菌胆固醇氧化酶。这些杂交质粒有效地转化了属于五个不同物种的双歧杆菌。开发了一组基于复制子pMB1的新型大肠杆菌-双歧杆菌穿梭载体（pLF5、pCLJ15、pSPEC1），其具有氯霉素、红霉素和壮观霉素抗性遗传决定因素，作为双歧杆菌的选择标记。质粒pTRE3是pLF5的衍生物，是最小的（2.8 kb）双歧杆菌载体，具有方便的多克隆位点，并且具有高结构稳定性和分离稳定性。
