Osaka University

Satoshi Harashima

Akio Kobayashi

Kiichi Fukui

Eiichiro Fukusaki

Yoshinobu Kaneko

Minetaka Sugiyama

Takefumi Sone

The present invention provides a simple method for splitting and loss of a chromosome in yeast. The method for modifying a chromosome in yeast includes preparing a linear chromosome splitting vector (1) having a target sequence (a), a marker gene sequence and (CA)sequence in this order; preparing a linear chromosome splitting vector (2) having a target sequence (b), a centromere sequence of a yeast chromosome and (CA)sequence in this order; and introducing the chromosome splitting vectors (1) and (2) into yeast. Herein, n is each independently an integer of 6 to 10. Although this chromosome splitting vector has a repetitive sequence of 5'-CCCCAA-3&#x2032;, it can be amplified specifically with PCR, so that a chromosome splitting vector can be prepared significantly simply and easily, compared with the conventional DNA splitting method. It seems that a yeast telomere sequence is bound to the (CA)sequence of the split chromosome generated by splitting with the chromosome splitting vector, and therefore the fragment can function as an intact chromosome.

Method for modifying chromosomes

A novel method for introduction of an exogenous substance or a physiologically active compound into cells is provided according to this invention. This method can realize introduction of an exogenous genetic substance or a physiologically active compound of large size with a large amount. Such substance is immobilized to beads of sphere fine particles having a particle size of 0.01 mm to 10 mm, and bio-active beads thus produced are introduced into cells. Bio-active beads comprising calcium alginate are particularly useful for the purpose of the present invention.

Method for introduction of an exogenous genetic substance or a physiologically active compound

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Osaka, Japan

Satoshi Harashima