摘要
Astructuralgene(750bp),whichcodesforatypeIribosomeinactivatingprotein,trichosanthin,hasbeendesignedaccordingtothecodonusageofhighlyexpressedgeneinE.coliandchemicallysynthesized.Inthesynthesizedgene,twenty-sevenuniquerestrictionsiteswereevenlydispersedwithanaveragedistancebetweentwoadjacentsiteslessthan50bptofacilitateasystematicinvestigationonstructure-functionalrelationshipofthisproteinbysite-directedmuta-genesis.Tosynthesizeit,thewholegenewasdividedintothreelargefragments(EP,PNandNH)whichwereassembledfromseveralchemicalsyntheticoligonucleotidesbyenzymaticmethod.TheassemblyofboththefragmentEPfromsixoligonucleotides(A-F)andthefragmentPNfromfouroligomers(G-J)wascatalyzedbyT4DNAligaseinusingthesinglestrandedDNAmethod[Chen,H.-B.etal,Nucl.AcidsRes.,18,871(1990)].AndfragmentNHwasformedfromthreeduplexesK,LandMbytheclassicaldoublestrandedDNAmethod.Finally,
出版日期
1995年04月14日（中国期刊网平台首次上网日期，不代表论文的发表时间）
