TERMINATOR SEQUENCE-CONTAINING REVERSE PRIMER FOR OVEREXPRESSION AND LINEAR DNA

摘要

Plasmid vectors have been widely used as a carrier of a DNA sequence capable of expressing a target RNA in cells. However, construction of these plasmid vectors requires technical skill and time. Thus, there has been required a more easy and quick method therefor. To solve this problem, a method using a linear DNA that has been amplified by the PCR method is examined. However, this method is disadvantageous in that RNA can be expressed in cells only at an extremely low level thereby. Under these circumstances, the present inventors attempted to develop an RNA expression method using a linear DNA which can be produced mainly by using the PCR method alone and which enables RNA expression at a high level. As the results of intensive studies on terminator sequences to be used in a linear DNA, the present inventors found a terminator sequence of the smallest unit which enables expression in a linear DNA equivalent to the expression with the use of a plasmid vector. A linear DNA comprising the aforesaid terminator sequence can be easily and quickly constructed and enables RNA expression at a higher level. The present invention has been completed based on these findings.