Batchikova N V, Al'tman I B, Lutsenko S V, Smirnov V A, Nazimov I V, Eshkind L G, Siniagina E A, Azhaev A V
Bioorg Khim. 1992 Jun;18(6):766-76.
Secretion vectors were constructed in which a synthetic gene of human epidermal growth factor (hEGF) joined with a gene coding for the leader peptide to one of the E. coli outer membrane major proteins (OmpF) is controlled by tac promoter. The increase of the hEGF yield was achieved by the multiplication of the gene copies. The hEGF in bacterial cells was secreted into periplasm. The recombinant protein was isolated by means of reverse phase chromatography as almost homogenous preparation (greater than 98%), the yield being 7 mg/l bacterial culture. The sequence of twenty-five N-terminal amino acid residues of the isolated hEGF coincided with that of the natural protein. The preparation proved to be biologically active.
构建了分泌载体，其中人表皮生长因子（hEGF）的合成基因与编码前导肽的基因连接到大肠杆菌外膜主要蛋白之一（OmpF），并由tac启动子控制。通过基因拷贝数的增加实现了hEGF产量的提高。细菌细胞中的hEGF被分泌到周质中。通过反相色谱法分离得到几乎纯的重组蛋白（纯度大于98%），产量为7mg/l细菌培养物。分离得到的hEGF的25个N端氨基酸残基序列与天然蛋白的序列一致。该制剂被证明具有生物活性。
