Chang C N, Rey M, Bochner B, Heyneker H, Gray G
Department of Cell Genetics, Genentech, Inc., South San Francisco, CA 94080.
Gene. 1987;55(2-3):189-96. doi: 10.1016/0378-1119(87)90279-4.
A gene encoding the mature form of human growth hormone (hGH) was fused to the secretion signal coding sequence of the Escherichia coli heat-stable enterotoxin II (STII). This hybrid gene was preceded by two Shine-Dalgarno sequences derived from the trp and STII-coding genes and was expressed in E. coli under the transcriptional control of the E. coli alkaline phosphatase (phoA) promoter. In low-phosphate growth media, cells synthesized about 15 to 25 micrograms of hGH/ml/1 A550 unit of cells. This represents 6 to 10% of total cellular protein. The majority of the hGH produced (more than 90%) was processed precisely and secreted into the periplasmic space. These results demonstrate that E. coli cells are able to synthesize and secrete high levels of this human protein using a prokaryotic signal sequence.
将编码人生长激素（hGH）成熟形式的基因与大肠杆菌热稳定肠毒素II（STII）的分泌信号编码序列融合。该杂种基因之前有两个源自色氨酸和STII编码基因的Shine-Dalgarno序列，并在大肠杆菌碱性磷酸酶（phoA）启动子的转录控制下在大肠杆菌中表达。在低磷酸盐生长培养基中，细胞每毫升每1个A550单位细胞合成约15至25微克hGH。这占细胞总蛋白的6%至10%。所产生的大部分hGH（超过90%）被精确加工并分泌到周质空间。这些结果表明，大肠杆菌细胞能够使用原核信号序列合成并分泌高水平的这种人类蛋白质。
