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| Expression of ITF Gene in Lacococcus lactis |
| BAI Cai-ming ,BI Yan-wei,YANG Xu,LI Zhi-hua,LI Jian-feng,XU Wei-ming |
| Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Kunming 650118, China |
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Abstract To develop an oral drug, ITF gene encoding ITF proein, was expressed in a live delivery vehicle lactococcus lactis. First, the ITF gene was cloned into the prokaryotic expressive vector pNICE:sec. Second, the recombinant vector pNICE:sec-ITF was transformated into Lactococcus lactis strain NZ9000 to express ITF protein. Then the recombinant ITF was induced to express and was identified by SDS-PAGE and Western blot.Rabbits are divided into blank control group,preparation group and therapeutic group which are respectively administrated wih PBS and pNICE:sec-ITF Lactococcus lactis. By grades of ulcer test whether administrated pNICE:sec-ITF Lactococcus lactis protects against HCl-induced gastric injury in rabbits. The results were described as follows. The ITF was amplified and cloned in the vector pNICE:sec successfully. The fusion protein (5.9kDa) was expressed in L. lactis by the induction of the nisin. The quantity of expression accounted for 5% of the total bacterial protein. Western bolt analysis confirmed that fusion protein could be recognized specially by Monoclonal Anti-human TFF3 Antibody. Preparation groups and therapeutic groups do good than control group. Prove that administrated pNICE:sec-ITF Lactococcus lactis is biologically active in an HCl-induced rabbit gastric mucosal injury model.
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Received: 04 March 2009
Published: 29 October 2009
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Corresponding Authors:
BAI Cai-Ming
E-mail: lijianfeng@imbcams.com.cn
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