A novel episomal shuttle vector for transformation of Cryptococcus neoformans with the ccdB gene as a positive selection marker in bacteria
P Mondon, YC Chang, A Varma… - FEMS microbiology …, 2000 - academic.oup.com
P Mondon, YC Chang, A Varma, KJ Kwon-Chung
FEMS microbiology letters, 2000•academic.oup.comWe report the engineering of a new shuttle vector featuring its episomal maintenance in
Cryptococcus neoformans and the lethal Escherichia coli ccdB gene for positive selection in
bacteria. Telomere-like sequences from C. neoformans and the STAB fragment confer
episomal maintenance to the vector (pPM8) upon transformation in C. neoformans. The
vector generated high transformation frequencies and each transformant was estimated to
harbor thirty copies of the plasmid. The plasmids recovered in E. coli from the C. neoformans …
Cryptococcus neoformans and the lethal Escherichia coli ccdB gene for positive selection in
bacteria. Telomere-like sequences from C. neoformans and the STAB fragment confer
episomal maintenance to the vector (pPM8) upon transformation in C. neoformans. The
vector generated high transformation frequencies and each transformant was estimated to
harbor thirty copies of the plasmid. The plasmids recovered in E. coli from the C. neoformans …
Abstract
We report the engineering of a new shuttle vector featuring its episomal maintenance in Cryptococcus neoformans and the lethal Escherichia coli ccdB gene for positive selection in bacteria. Telomere-like sequences from C. neoformans and the STAB fragment confer episomal maintenance to the vector (pPM8) upon transformation in C. neoformans. The vector generated high transformation frequencies and each transformant was estimated to harbor thirty copies of the plasmid. The plasmids recovered in E. coli from the C. neoformans transformants showed no evidence of rearrangement. This construct will be very useful for cloning and studying the regulation of genes in C. neoformans.
Oxford University Press