| 摘要 |
PURPOSE:To provide a DNA fragment composed of at least a part of a specific nucleotide sequence or of a base sequence equivalent to the nucleotide sequence, having promoter activity and useful for gene recombination technique, etc. CONSTITUTION:(A) Whole DNA of raw leaf of barley is nicked with a restriction enzyme BamHI to obtain a DNA fragment of 243 bp. (B) Separately, an E.coli plasmid pKK232-8 containing a restriction enzyme BamHI nicking site; structural gene CAT of chloramphenicol acetyltransferase; ampicillin-resistant gene Amp originated from pBR322, etc., is completely decomposed with BamHI and dephosphorylated to obtain a treated DNA. The components A and B are linked together using a T4 DNA ligase and E.coil C600 is transformed with the obtained recombinant plasmid and selected in a selection medium to obtain a linked plasmid pBT2402 containing a DNA fragment having strongest promoter activity. The plasmid is nicked with BamHI to obtain a promoter containing a DNA fragment of formula composed of 243 base pairs. |
