Zhou Yuxun, Cao Wei, Wang Jinzhi, Ma Yushu, Wei Dongzhi
State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, P.R.
Protein Pept Lett. 2005 May;12(4):349-55. doi: 10.2174/0929866053765671.
Adenoregulin is a 33 amino acid antibiotic peptide who belongs to dermaseptin family which is the first vertebrate family to show lethal effects against filamentous fungi, as well as a broad spectrum of pathogenic microorganisms. Synthetic adenoregulin gene was cloned in 2, 4 and 6 tandem repeats and subcloned in pET32a and pET22b vectors. Recombinant plasmids were transformed into E. coli BL21(DE3), Fusion proteins of Trx-ADR1, Trx-ADR2 and Trx-ADR4 could be expressed after the hosts were induced by IPTG, but the expression level decreased dramatically with the number of tandem repeats increased. ADR1, ADR4 and ADR6 could not be expressed by E. coli without carrier proteins. But for Pichia pastoris GS115, ADR1 and ADR6 in the fermentation broth of the hosts could be detected by ELISA, and the bactericidal activities could also be observed.
腺调节素是一种由33个氨基酸组成的抗菌肽，属于皮栖菌素家族，是首个对丝状真菌以及多种致病微生物具有致死作用的脊椎动物家族。合成的腺调节素基因以2、4和6个串联重复序列进行克隆，并亚克隆到pET32a和pET22b载体中。重组质粒被转化到大肠杆菌BL21(DE3)中，经IPTG诱导宿主后，Trx-ADR1、Trx-ADR2和Trx-ADR4融合蛋白能够表达，但随着串联重复序列数量的增加，表达水平显著下降。没有载体蛋白时，大肠杆菌无法表达ADR1、ADR4和ADR6。但对于毕赤酵母GS115，通过ELISA可检测到宿主发酵液中的ADR1和ADR6，并且还能观察到杀菌活性。
