Ghorpade A, Garg L C
Gene Regulation Laboratory, National Institute of Immunology, New Delhi, India.
FEBS Lett. 1993 Sep 6;330(1):61-5. doi: 10.1016/0014-5793(93)80920-p.
The heat-labile enterotoxin chain B (LTB) signal sequence was used for the processing and export of human growth hormone (hGH). The protein was completely processed and exported across the cell membrane to accumulate in the periplasmic space in Escherichia coli. The human growth hormone cDNA was cloned as a PCR amplified fragment under the control of tac promoter and translationally fused to the LTB signal sequence. The rate of processing of hGH under the control of the LTB signal sequence was equal to or more than the rate of induction of expression, indicating efficient processing. The receptor binding activity of the processed periplasmic protein was established in a radio receptor assay.
热不稳定肠毒素B链(LTB)信号序列用于人生长激素(hGH)的加工与分泌。该蛋白在大肠杆菌中得到完全加工,并穿过细胞膜分泌至周质空间进行积累。人生长激素cDNA作为PCR扩增片段,在tac启动子的控制下进行克隆,并与LTB信号序列进行翻译融合。在LTB信号序列控制下,hGH的加工速率等于或高于表达诱导速率,表明加工效率较高。通过放射性受体分析确定了加工后的周质蛋白的受体结合活性。