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首页> 外国专利> Recombinant plasmid DNA pER-TA1 GyrA-AcSer, encoding the serine acetyltransferase, the ability of IN VIVO acetylated N-terminal serine Desacetylthymosin ALPHA 1 and the hybrid protein capable of autocatalytic cleavage with formation Thymosin Alpha 1 HUMAN STRAIN Eschrichia coli C3030 / pER-TA1GyRA-AcSer PRODUCER said proteins and methods for producing genetically engineered thymosin Alpha 1 PERSON

Recombinant plasmid DNA pER-TA1 GyrA-AcSer, encoding the serine acetyltransferase, the ability of IN VIVO acetylated N-terminal serine Desacetylthymosin ALPHA 1 and the hybrid protein capable of autocatalytic cleavage with formation Thymosin Alpha 1 HUMAN STRAIN Eschrichia coli C3030 / pER-TA1GyRA-AcSer PRODUCER said proteins and methods for producing genetically engineered thymosin Alpha 1 PERSON

机译：重组质粒DNA pER-TA1 GyrA-AcSer，编码丝氨酸乙酰基转移酶，IN VIVO乙酰化N端丝氨酸脱乙酰胸腺素ALPHA 1的能力以及能够自动催化裂解并形成胸腺素α1人类应变埃希氏大肠杆菌C3030 -AcSer PRODUCER表示用于生产基因工程胸腺素Alpha 1 PERSON的蛋白质和方法

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1. Recombinant DNA plasmid pER - TA1GyrA - AcSer, comprising polycistronic construct for biosynthesis two proteins simultaneously - a hybrid polypeptide comprising human thymosin α1 and intein and enzyme E.coli - serine acetyltransferase in cells of Escherichia coli; NdeI / SapI - fragment of plasmid DNA pTWIN-1; NdeI / SapI - DNA fragment containing these sites adapted to the gene sequence of recombinant human thymosin α1 and comprising, as a genetic marker β-lactamase gene that determines the resistance of transformed plasmid pER - TA1GyrA - AcSer E.coli cells to penicillin antibiotics; unique restriction endonuclease recognition sites located at a distance next to the left from the site BamHI - Pstl - 575 bp, NdeI -: 1448 bp, XbaI - 1487 bp, EcoRV - 3520 bp, HpaI - 3576 p.o.2. Escherichia coli Strain S3030 / pER - TA1GyrA - AcSer, producing a hybrid polypeptide comprising human α1 Desacetylthymosin and intein and serine acetyltransferase obtained by transforming Escherichia coli cells of the strain S3030 recombinant DNA plasmid pER - TA1GyrA - AcSer of claim 1.3.. A method for producing recombinant thymosin α1 human comprising transformation of strain Escherichia coli S3030 plasmid DNA pER - TA1GyrA -. AcSer, according to claim 1, culturing the producing strain Escherichia coli S3030 / pER - TA1GyrA -. AcSer according to claim 2, separation TA1GyrA fusion protein in the soluble form following cell disruption using ultrasonic disintegrator in a buffer solution (50 mM Tris / HCl, 10 mM EDTA, 1 mM PMSF), adsorbing the fusion protein on a column of chitinous sorbent (NEB, England), inducing its autocatalytic cleavage washing chitin sorbent buffer containing equilibrated in buffer comprises
机译：1.重组DNA质粒pER-TA1GyrA-AcSer，其包含用于同时生物合成两种蛋白质的多顺反子构建体-在人大肠杆菌细胞中包含人胸腺素α1和内含肽以及大肠杆菌酶-丝氨酸乙酰基转移酶的杂合多肽; NdeI / SapI-质粒DNA pTWIN-1的片段; NdeI / SapI-DNA片段，包含与重组人胸腺素α1的基因序列相适应的这些位点，并且包含作为遗传标记的β-内酰胺酶基因，其确定了转化的质粒pER-TA1GyrA-AcSer大肠杆菌细胞对青霉素抗生素的抗性;唯一限制性内切核酸酶识别位点，位于位点BamHI-Pstl-575 bp，NdeI-：1448 bp，XbaI-1487 bp，EcoRV-3520 bp，HpaI-3576 p.o. 14.一种大肠杆菌菌株S3030 / pER-TA1GyrA-AcSer，产生包含人α1去乙酰胸腺素和内含肽以及丝氨酸乙酰基转移酶的杂合多肽，所述杂合多肽是通过转化权利要求1.3所述的菌株S3030重组DNA质粒pER-TA1GyrA-AcSer获得的。产生重组胸腺素α1人，其包括转化大肠杆菌S3030菌株质粒DNA pER-TA1GyrA-。 2.根据权利要求1所述的AcSer，培养生产菌株大肠杆菌S3030 / pER-TA1GyrA-。 3.根据权利要求2所述的AcSer，在缓冲液（50mM Tris / HCl，10mM EDTA，1mM PMSF）中使用超声崩解剂破坏细胞后，以可溶性形式分离TA1GyrA融合蛋白，将融合蛋白吸附在几丁质吸附剂柱上（NEB，英格兰），诱导其自动催化裂解洗涤包含在缓冲液中平衡的几丁质吸附剂缓冲液，包括

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