Genetic Approaches to the Detection of Contaminants in Escherichia coli Fermentations

详细信息    查看全文

• 作者：James R. Swartz and and Nancy McFarland
• 刊名：Biotechnology Progress
• 出版年：1998
• 出版时间：February 1998
• 年：1998
• 卷：14
• 期：1
• 页码：88 - 91
• 全文大小：40K
• 年卷期：v.14,no.1(February 1998)
• ISSN：1520-6033

文摘

In facilities which cultivate more than one rDNA organism, contaminationby the samespecies is difficult to detect. Since the same species may beproducing a differentproduct in an adjacent fermentor or in the same vessel in a subsequentprocedure,the possibility of cross-product contamination must be considered.Here we describea simple, sensitive, and reliable technique for the detection ofsame-species contamination. The assay uses negative genetic markers such as the inabilityto use acarbohydrate, e.g., ribose. When the facility is managed to usethe ribose marker foronly one product, this culture can be plated on ribose minimal mediumto allow rapidand sensitive detection of contaminants. If the facility is usedfor several rDNAproducts, multiple carbohydrate markers per strain can be used so thata limitednumber of markers can differentiate among larger host collections.The approach wasdeveloped and tested using Escherichia coli as the hostorganism. If hosts withauxotrophies, e.g., for amino acids, are used in the facility, theplate medium can besupplemented. When this technique is combined with existingmethods for detectingdifferent species and bacteriophage contamination, all three classes ofbiologicalcontamination can be detected.