HUMAN PROTEIN C GENE HAVING NOVEL BASE SEQUENCE

摘要

PURPOSE:To provide a human protein C gene free from intron part, containing a number of newly introduced restriction enzyme recognition sites, composed of partial substitute for synthetic DNA fragment and easily treatable by protein engineering operation. CONSTITUTION:The base sequence of a human protein C protein is varied by using a known base sequence as a base for about 2/3 of the protein from N-terminal while keeping the coded amino acids unchanged. The above region is supposed to be important in the expression and function of human protein C protein. The human protein C is divided by this process into 30 small DNA fragments each having a length of 33-80 nucleotides and designed to have restriction enzyme recognition sites at both ends. The divided small fragments are individually synthesized, subsequently linked with each other and cloned to obtain a synthetic part. The remaining part corresponding to 1/3 of the protein and originated from chromosome gene is linked with the above synthetic part using BamHI-HaeII fragments of the 8th exon which is the longest exon of the chromosome gene of the protein C. The objective gene coding 1389 human protein C and free from intron part can be produced by this process.