EXPRESSION OF HUMAN IL-2 GENE IN ESCHERICHIA COLI

  • Trần Ngọc Tân

Abstract

Interleukin-2 (IL-2) is historically known as a T-cell growth factor. It is a centtal regulator in the immune system and mediates its effects. Administtation of high or intermediate doses of exogenous IL-2 has been used as an anti-tumor tteatment in humans, especially for renal and melanoma. Therefore, in  1992, the high-dose of recombinant IL-2 was approved by the Food and Dmg Administtation  (FDA) to use for  cancer tteatment. In order to develop recombinant IL-2 in Vietaam, we established a new strategy to express IL-2 protein from E. coli. In this sttategy, IL-2 which was not fused with any other protein, was expressed as inclusion bodies. The

recombinant protein was recovered efficiently  by the method of denatured purification  to separate recombinant IL-2 from other unexpected soluble proteins expressed in E. coli cells. In this report, the result of human il-2mn gene expression was shown. Based on the available pET32-Trx-IL2MN plasmid, the gene sequence was easy to  amplify  by  PCR with  specific  primer  pair  and  cloned  into pET22b(-i-) vector  in E.  coli BL21  cell.  After expression in LB medium, the experiment result showed that the optimal conditions to express recombinant IL-2 should be at 37°C with  1  mM final concenttation of inducer isopropyl-beta-D-thiogalactopyranoside (IPTG)

and the point to start induction at high cell density (OD ^oo from 0.9 to 1.2). This resuh also demonsttated that most expressed recombinant IL-2 was in the form of inclusion bodies as the purifying  sttategy expected.

Tác giả

Trần Ngọc Tân
điểm /   đánh giá
Published
2011-11-17