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首页> 外国专利> Production of purified somatostatin in E. coli cells comprises expression in the cells of the coding gene cloned in the vector pGRV1, with digestion by cyanogen bromide for chromatographic purification

Production of purified somatostatin in E. coli cells comprises expression in the cells of the coding gene cloned in the vector pGRV1, with digestion by cyanogen bromide for chromatographic purification

机译：在大肠杆菌细胞中生产纯化的生长抑素包括在载体pGRV1中克隆的编码基因在细胞中表达，并用溴化氰消化以进行色谱纯化

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Producing and purifying somatostatin in Escherichia colicells using isopropyl-beta-D-thiogalactopyranoside (IPTG) inducer molecule, based on the expression of a gene encoding for somatostatin pGRV1 cloned in a plasmid, comprises: (a) cloning the somatostatin gene in the plasmid pGRV1 to obtain pSOMA vector; (b) transforming in E. colistrain BL21-DE3 with plasmid pSOMA, to obtain the strain CECT5840; (c) fermenting strain CECT5840; and (d) extracting protein, permeabilizing inclusion bodies, and chromatographic purification to obtain somatostatin. Producing and purifying somatostatin in E. colicells using isopropyl-beta-D-thiogalactopyranoside (IPTG) inducer molecule, based on the expression of a gene encoding for somatostatin pGRV1 cloned in a plasmid, comprises: (a) designing a gene encoding human somatostatin comprising a fully defined 54 bp sequence (SEQ ID NO. 2) given in the specification; (b) constructing plasmid pGRV1 comprising a fully defined 6343 bp sequence (SEQ ID NO. 8) given in the specification; (c) cloning the somatostatin gene in the plasmid pGRV1 to obtain pSOMA vector; (d) transforming in E. colistrain BL21-DE3 with plasmid pSOMA, to obtain the strain CECT5840; (e) fermenting strain CECT5840 in SL media under controlled conditions; and (f) extracting protein, permeabilizing inclusion bodies, and chromatographic purification to obtain somatostatin.
机译：使用异丙基-β-D-硫代半乳糖吡喃糖苷（IPTG）诱导剂分子在大肠杆菌细胞中生产和纯化生长抑素，基于在质粒中克隆的编码生长抑素pGRV1的基因的表达，包括：（a）在质粒pGRV1中克隆生长抑素基因获得pSOMA载体; （b）用质粒pSOMA转化大肠杆菌BL21-DE3，得到菌株CECT5840。 （c）发酵菌株CECT5840; （d）提取蛋白质，使包涵体通透，并进行色谱纯化，以获得生长抑素。使用异丙基-β-D-硫代半乳糖吡喃糖苷（IPTG）诱导剂分子在大肠杆菌细胞中生产和纯化生长抑素，基于克隆在质粒中的生长抑素pGRV1编码基因的表达，该方法包括：（a）设计编码人生长抑素的基因，该基因包括说明书中给出的完整定义的54 bp序列（SEQ ID NO.2）; （b）构建包含说明书中给出的完整定义的6343bp序列（SEQ ID NO.8）的质粒pGRV1; （c）克隆质粒pGRV1中的生长抑素基因以获得pSOMA载体; （d）用质粒pSOMA转化大肠杆菌BL21-DE3，得到菌株CECT5840。 （e）在受控条件下在SL培养基中发酵菌株CECT5840; （f）提取蛋白质，使包涵体通透，并进行色谱纯化，以获得生长抑素。

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