高含量人体必需氨基酸基因的化学全合成  

CHEMICAL SYNTHESIS OF A GENE CODING FOR HIGH ESSENTIAL AMINO ACIDS OF HUMANS

作　　者：张晓东[1] 杨迪[2] 汪训明[2] 闵永洁 谢毅[2] 王鄂生[1] 王启松[2] 

机构地区：[1]武汉大学生物学系 [2]复旦大学遗传学研究所

出　　处：《复旦学报（自然科学版）》1990年第2期189-197,共9页Journal of Fudan University：Natural Science

摘　　要：采用固相亚磷酰胺法合成了编码高含量人体必需氨基酸蛋白质的基因。该基因有300个碱基对,共编码94个氨基酸,其中92%以上为人体必需氨基酸。从该基因的两个方向均可被翻译成具有人体必需氨基酸的蛋白质。该基因选用了植物所偏爱的密码子,并间隔地插入了编码赖氨酸的密码子,使之所编码的蛋白质可被胰蛋白酶消化。A gene coding for high essential amino acids of humans was synthesized byusing the solid phosphoramide method. The gene is 300 bp in length and encodes94 amino acids except two restriction sites at the end. More than 92% of the a.a.are essential for humans. When the gone is fused to the plant gene promotor in ei-ther direction, the expressed gone product will be a protein of high essential aminoacids. The plant preferred codons were chosen for the gone. Codons encodinglysines were inserted into this gene at interval. In order to modify and seperatethe two large fragments to be assembled, the EcoR I, EcoR I and BamH I, HindIII restriction sites were designed at 5′end, 3′end and the 141 bp site of the gone,respectively. Two large fragments were assembled by the methed of 'Three StepsSeperating Annealing Assembling' and inserted into M13mp18RF DNA sepera-tely. The identity of the two fragments was checked by DNA sequencing. Aftertwo DNA fragments were isolated, purified ligated, and inserted into M13mp18RF DNA and trasformed. The identity of the gone was checked by enzymaticdigestion and DNA sequencing.

关 键 词：氨基酸 基因 合成 亚磷酰胺法 人体 

分 类 号：Q781[生物学—分子生物学]