High frequency of yeast transformation by plasmids carrying part or entire 2-micron yeast plasmid. |
| |
Authors: | C Gerbaud P Fournier H Blanc M Aigle H Heslot M Guerineau |
| |
Institution: | 1. Laboratoire de Biochimie Enzymologie, Institut Gustave-Roussy, 94800 Villejuif, France;2. Laboratoire de Génétique, Institut National Agronomique, 75005, Paris, France;3. Laboratoire de Génétique Physiologique, Institut de Biologie Moléculaire et Cellulaire du CNRS, 67084 Strasbourg, France;4. Centre de Génétique Moléculaire du CNRS, 91190 Gif-sur-Yvette France |
| |
Abstract: | By using two chimeric plasmids containing yeast ura3 gene and 2-micron yeast DNA linked to the bacterial plasmid pCR1, yeast transformation of a high frequency has been achieved. The first plasmid is such that the 2-micron DNA part, in which the ura3 gene is incorporated, can be removed in one step and thus the 2-micron-ura3 sequence can be considered as a "transposable" block. In contrast, the second one bears the entire 2-micron plasmid and the ura3 gene is inserted in the bacterial plasmid part. As shown through hybridization experiments and genetic studies, the ura3 gene was maintained as a cytoplasmic element. Plasmids recovered from the yeast transformants were used to transform Escherichia coli. Their analysis by EcoRI showed that in many cases the vector had recombined with the endogenous 2-micron DNA of the recipient strain. The specific activity of orotidine 5'-monophosphate decarboxylase (coded by ura3) in yeast transformants was 10- to 30-fold higher than in the wild type. |
| |
Keywords: | Chimeric plasmids 2-μm DNA pCR1 recombinant DNA |
本文献已被 ScienceDirect 等数据库收录! |
|