| 摘要 |
PURPOSE:To obtain a novel anti-tumor polypeptide deficient of 11-amino acid of amino terminal of TNF, by linking a synthetic oligonucleotide with a proper promoter and introducing the obtained recombinant plasmid into E.coli. CONSTITUTION:A human TNF gene having translation-initiation codon, termination codon and a proper restriction enzyme site is designed. Said gene is divided into 3 blocks by using 17 synthetic oligonucleotides and cloned to vector pBR322. Fragment of each block (ClaI-SalI, SalI-PvuII, PvuII-HindIII) and a ClaI-HindIII fragment of a plasmid PYS31N having trp promoter are linked together and introduced into E.coli to obtain pTNF401NN. The HapII-HindIII fragment of said plasmid is linked with ClaI-HindIII fragment of pYS31N and further a synthetic oligonucleotide and the product is introduced into a host. The objective polypeptide having the amino acid sequence of formula can be produced by using said host cell. |
