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Plasmids for production of met-asp-gln bovine growth hormone, hosts containing the plasmids, products manufactured thereby and related methods

机译:用于生产met-asp-gln牛生长激素的质粒,含有该质粒的宿主,由此生产的产品以及相关方法

摘要

An improved vector upon introduction into a suitable bacterial host containing the thermolabile repressor C.sub.I renders the host cell capable, upon increasing the temperature of the host cell to a temperature at which the repressor is destroyed, of effecting expression of a desired gene inserted into the vector and production of polypeptide encoded by the gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: a DNA sequence which contains the promoter and operator P.sub.L O.sub.L from lambda bacteriophage; the N utilization site for binding antiterminator N protein produced by the host cell; a DNA sequence which contains a ribosomal binding site for rendering the mRNA of the desired gene capable of binding to ribosomes within the host cell; an ATG initiation codon or a DNA sequence which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a restriction enzyme site for inserting the desired gene into the vector in phase with the ATG initiation codon; and additionally a DNA sequence which contains an origin of replication from a bacterial plasmid capable of autonomous replication in the host cell and a DNA sequence which contains a gene associated with a selectable or identifiable trait which is manifested when the vector is present in the host cell.
机译:引入含有热不稳定阻遏物C.I的合适细菌宿主中的改进载体使宿主细胞能够在将宿主细胞的温度升高至破坏阻遏物的温度时影响所需基因的表达。插入载体中并产生该基因编码的多肽。载体是双链DNA分子,其以5'至3'顺序包括:DNA序列,其包含来自λ噬菌体的启动子和操纵子PLOL;结合宿主细胞产生的抗终止子N蛋白的N利用位点;含有核糖体结合位点的DNA序列,用于使所需基因的mRNA能够结合宿主细胞内的核糖体。 ATG起始密码子或将所需基因插入载体后转化为ATG起始密码子的DNA序列;用于将所需基因与ATG起始密码子同相插入载体的限制性酶切位点;另外,该DNA序列含有一个能在宿主细胞中自主复制的细菌质粒的复制起点,以及一个DNA序列,该DNA序列含有一个与可选择或可鉴定的性状相关的基因,该基因在载体存在于宿主细胞中时即表现出来。 。

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