首页> 外国专利> RECOMBINANT PLASMID DNA pET32M/mTrx-rhIL-11 CODING HUMAN INTERLEUKIN-11, METHOD OF PRODUCING AND Escherichia coli BL21 (DE3)/pET32M/mTrx-rhIL-11 STRAIN - PRODUCER OF RECOMBINANT INTERLEUKIN-11

RECOMBINANT PLASMID DNA pET32M/mTrx-rhIL-11 CODING HUMAN INTERLEUKIN-11, METHOD OF PRODUCING AND Escherichia coli BL21 (DE3)/pET32M/mTrx-rhIL-11 STRAIN - PRODUCER OF RECOMBINANT INTERLEUKIN-11

机译:重组质粒DNA pET32M / mTrx-rhIL-11编码人类白细胞介素11,生产方法和大肠杆菌BL21(DE3)/ pET32M / mTrx-rhIL-11菌株-重组白细胞介素11的生产者

摘要

FIELD: medicine.;SUBSTANCE: invention can be used for producing recombinant human interleukin-11 (rIL-11). The recombinant plasmid DNA pET32M/mTrx-rhIL-11 is produced by a method which involves synthesis of four fragments of a recombinant human interleukin-11 gene from oligonucleotide primers, cloning of each fragment in a plasmid vector pGEM5z restricted by EcoRV endonuclease, transformation of each of 4 plasmids of E.coli cells of strain DH5a, selection of clones coding the related fragments of the recombinant human interleukin-11 gene, selection of plasmids with the recombinant human interleukin-11 gene without mutations, combination of all fragments of the recombinant human interleukin-11 gene in the plasmid vector pUC19, construction of the vector pET32M of the plasmid pET32b, recloning of the recombinant human interleukin-11 gene in an expression vector pET32M, coding thioredoxin 1 E.coli with substitution of Asn84/Gln. The produced DNA is used to transform the E.coli BL21 (DE3) strain cells to produce the E.coli BL21 (DE3)/pET32M/mTrx-rhIL-11 strain that is a producer of recombinant human interleukin-11 as an ingredient of soluble fused protein mTrx-rhlL-11.;EFFECT: effective production of recombinant interleukin-11 in Escherichia coli cells.;3 cl, 5 dwg, 1 tbl, 4 ex
机译:领域:药物;发明:本发明可用于生产重组人白介素11(rIL-11)。重组质粒DNA pET32M / mTrx-rhIL-11的制备方法涉及从寡核苷酸引物合成重组人白细胞介素11基因的四个片段,将每个片段克隆到受EcoRV核酸内切酶限制的质粒载体pGEM5z中,菌株DH5a的大肠杆菌细胞的4个质粒中的每一个,选择编码重组人白细胞介素11基因相关片段的克隆,选择具有重组人白细胞介素11基因且无突变的质粒,重组体所有片段的组合质粒载体pUC19中的人白细胞介素11基因,质粒pET32b的载体pET32M的构建,表达载体pET32M中重组人白细胞介素11基因的克隆,编码硫氧还蛋白1大肠杆菌并被Asn84 / Gln取代。产生的DNA用于转化大肠杆菌BL21(DE3)菌株细胞,以生产大肠杆菌BL21(DE3)/ pET32M / mTrx-rhIL-11菌株,该菌株是重组人白介素11的生产者,是大肠杆菌的一种成分。可溶性融合蛋白mTrx-rhlL-11 。;效果:在大肠杆菌细胞中有效产生重组白介素11; 3 cl,5 dwg,1 tbl,4 ex

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