A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli.

A method, using LiAc to yield competent cells, is described that increased the efficiency of genetic transformation of intact cells of Saccharomyces cerevisiae to more than 1 × 105 transformants per microgram of vector DNA and to 1.5% transformants per viable cell. The use of single stranded, or heat denaturated double stranded, nucleic acids as carrier resulted in about a 100 fold higher frequency of transformation with plasmids containing the 2μm origin of replication. Single stranded DNA seems to be responsible for the effect since M13 single stranded DNA, as well as RNA, was effective. Boiled carrier DNA did not yield any increased transformation efficiency using spheroplast formation to induce DNA uptake, indicating a difference in the mechanism of transformation with the two methods.

High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier.

mRNA stability in mammalian cells.

Molecular biologists are increasingly faced with the problem of assigning a function to genes that have been cloned. A new approach to this problem involves the manipulation of the cloned gene to create what are known as 'dominant negative' mutations. These encode mutant polypeptides that when overexpressed disrupt the activity of the wild-type gene. There are many precedents for this kind of behaviour in the literature--some oncogenes might be examples of naturally occurring dominant negative mutations.

Functional inactivation of genes by dominant negative mutations

http://biscweb.uark.edu/ipinto/biol5334/5.pdf

Douglas Meeks-Wagner

Papers

Normal stoichiometry of histone dimer sets is necessary for high fidelity of mitotic chromosome transmission.

A rapid, efficient method for isolating DNA from yeast.

Isolation of two genes that affect mitotic chromosome transmission in S. cerevisiae