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    • 6. 发明授权
    • Processes for the preparation of foreign proteins in streptomycetes
    • 在链霉菌中制备外源蛋白的方法
    • US5426036A
    • 1995-06-20
    • US30731
    • 1993-03-12
    • Klaus-Peter KollerGunther RiessEugen UhlmannHolger Wallmeier
    • Klaus-Peter KollerGunther RiessEugen UhlmannHolger Wallmeier
    • C07K14/36C07K14/62C12N15/76C12N15/62
    • C07K14/62C07K14/36C12N15/76C07K2319/02C07K2319/75
    • The tendamistat gene can be used for the construction of fused genes with which fusion proteins are expressed and excreted in Streptomycetes host cells. The tendamistat portion can be modified, in particular it can be C-terminal shortened. When a gene for a shortened proinsulin in which the insulin B chain is linked to the A chain only via Lys or Lys-Lys is coupled to the tendamistat gene, this gene construction is introduced into an expression vector, and the latter is used to transform a Streptomycetes host cell, there is expression and secretion of the corresponding fusion protein. The fusion protein can easily be cleaved to give insulin precursors because of correctly established disulfide bonds. Genetic structures which code for the signal sequence and about the first ten amino acids of tendamistat as well as a desired protein are expressed in streptomyces host cells with a high yield, and the fusion proteins are secreted into the medium.
    • 该趋化因子基因可用于构建融合蛋白的融合蛋白,其在链霉菌宿主细胞中表达和排泄。 可以修改卷曲部分,特别是可以缩短C端。 当胰岛素B链仅通过Lys或Lys-Lys与A链连接的缩短的胰岛素原的基因与趋化因子基因偶联时,将该基因构建引入表达载体中,并将​​后者用于转化 链霉菌宿主细胞,有相应融合蛋白的表达和分泌。 由于正确建立的二硫键,融合蛋白可以容易地被切割以产生胰岛素前体。 编码信号序列和约10个氨基酸的趋化因子以及所需蛋白质的遗传结构以高产量在链霉菌宿主细胞中表达,融合蛋白分泌到培养基中。