DESIGNING AND CONSTRUCTION OF TRANSCRIPTION TEMPLATE FOR CELL-FREE PROTEIN SYNTHESIS, AND DILUTION BATCH WHEAT EMBRYO CELL-FREE PROTEIN SYNTHESIS METHOD USING THE SAME

摘要

To design and construct a transcription template for synthesizing a cell-freeprotein which is commonly usable, has a high translation template activity andcan be easily constructed, a polynucleotide comprising a base sequenceinherent to an individual vector capable of forming a complementary strandwith a base sequence existing between the transcriptional terminator sequenceof a marker gene (for example, a drug-tolerance gene) of the vector and Ori isprovided as a 3'-terminal PCR primer, while two types of primers satisfyingthe requirement of not undergoing transcription from a DNA constructed byusing a single primer alone, involving a polynucleotide having a sequencecomplementary to a base sequence containing a part of the promoter functionsite from the 5'-terminus of a promoter and another polynucleotide having asequence complementary to a base sequence containing a part of the RNApolymerase recognition site from the 3'-terminus of the promoter are providedas primers for 5'-terminal PCR. Further, a method of constructing atranscription template, etc. by using them is provided.