摘要
用限制性内切酶将人尿激酶原cDNA酶切成一系列基因片段 ,并分别在大肠杆菌中表达。发现其中 1个富含大肠杆菌稀有密码子AGG(精氨酸 )的片段表达量低 ,成为人尿激酶原cD NA在E .coli中高效表达的限制因素。将富含AGG(精氨酸 )的片段在E .coliBL2 1 CodonPlusTM RIL中表达 ,通过该菌株引入dnaY基因 (即tRNAagg/aga(Arg) )使该片段的表达量提高了 10倍。最后用同样的方法提高全长人尿激酶原cDNA在大肠杆菌中的表达量 ,使其表达量达到全菌蛋白的5%。
Studies were made on the enhancement of expression of human pro\|Urokinase(Pro\|UK) cDNA in \%Eschevichia coli.\% By means of PCR,the signal peptide DNA sequence was deleted.For study of the limited factors in expression of pro\|Urokinase,the pro\|UK gene was divided into three fragments,and they were expressed in E.coli BL21(DE3),The expression of middle fragment is very low,because there are several rare codon AGG (Arg) in it.Using a new bacteria E.coli BL21\|CodonPlus TM RIL to introduce dnaY gene coding tRNA agg/aga (Arg) to recognize the rare codon AGG,the expression level of the middle fragment was enhanced to 10%～20% of total cell protein and the expression of the intact pro\|UK was enhanced to 5% by more than 10 folds.
出处
《北京大学学报（自然科学版）》
CAS
CSCD
北大核心
2000年第6期802-807,共6页
Acta Scientiarum Naturalium Universitatis Pekinensis
关键词
尿激酶原
大肠杆菌
稀有密码子
dnaY基因
表达
Pro\|Urokinase
\%Eschevichia coli\%
rare codon
BL21\|CodonPlus TM RIL
dnaY gene
