2006 Fiscal Year Final Research Report Summary
Functional analyais of defense molecule using transgenic mice
| Project/Area Number |
16390369
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Digestive surgery
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| Research Institution | Tohoku University |
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Principal Investigator |
FUKUSHIMA Kouhei Tohoku University, Tohoku Univ.hospital, Lecture (20271900)
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| Co-Investigator(Kenkyū-buntansha) |
SASAKI Iwao Tohoku Univ., Graduate School of Medicine, Professor (60125557)
SHIBATA Chikashi Tohoku University, Tohoku Univ.Hospital, Assistant Professor (30270804)
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| Project Period (FY) |
2004 – 2006
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| Keywords | mucosal defense / transgenic mice / epithelial cells / ulcerative colitis / intestinal flora / total proctocolectomy |
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| Research Abstract |
The aims of the present study were to analyze molecular function of resistin-like molecule β (RELM-β) and deleted in malignant brain tumors 1(DMBT-1), which had been identified in colonic epithelial cells by bacterial reconstitution of germ-free mice.
We established RELM-β transgenic mice using pCAG vector. After a long time and started to investigate expression of the trannsgene. Its expression in the colon was unexpectedly weak due to unknown reasons. In addition, there was no significant difference between control and TG mice after challenge of dextran sulfate sodium, which are well accepted as an inducer of colitis in mice.
In vitro experiments, we found antibacterial effects of RELM-β and still try to confirm this function. Recombinant RELM-β bound to the cell surface of Staphylococcus aureus and destroyed its membrane. This protein is present as dimmer in stools.
On the other hand, we struggled to clone full-length DMBT-1 and make its recombinant protein and finally failed. The recent report suggests that cloning of DMBT-1 requires exchange of nucleotides from original to modified sequences (Caroline End, et al. Protein expression and purification 41, 275 (2005).
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