PURPOSE: To facilitate production of study of human lymphotoxin by providing one place of recognition site of 9 kinds of restriction enzymes to recognize 6 base pairs of palindromes, eliminating nucleotide sequence of specific part and providing specific recognition sequence.
CONSTITUTION: Nucleotide sequence of human lymphotoxin is modified in the following way to give a synthetic gene. (1) A methionine codon is added to the upper stream of leucine codon, two nonsense codons are added in series to 3'-end and further ClaI to 5'-end and SalI recognition sequence to 3'-end in order to efficiently carry out translation initiation. (2) One place is provided with recognition site of restriction enzyme BssHII, EcoRI, KpnI, HinaIII, XhoI, SacI, SmaI, HpaI and BamHI to recognize 6 pairs of palindromes. (3) Chain length of 100 nucleotides is made into one block and direct repeat in the block is eliminated. (4) Palindromes sequence exceeding 6 base pairs are not made to exist in any of continuous 30 nucleotides.
MIURA KAZUNOBU
UESUGI HARUICHI
NISHIKAWA SATOSHI
MATSUO TOKUYUKI
