| Budget Amount *help |
¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2001: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2000: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Research Abstract |
The toughness is one of the most important elements, which define the commercial value of raw meat of fish. The degradation of extracellulr matrix is thought to be a cause of post-mortem tenderization of fish meat. Our previous study suggested that this tenderization is caused mainly by the metalloproteinases. Especially, matrix metalloproteinases (MMPs) are thought to be the most possible candidates for post-mortem tenderization of fish meat.
To elongate the toughness of fish meat during chilled storage, we proposed new technique mentioned below. There are endogenous proteinase inhibitors specific for MMPs. They are designated as tissue inhibitore of metalloproteinases (TIMPs). We attempt to elevate the quantity of TIMPs in fish meat during alive. At present, gene manipulated food has not get public acceptance yet So, we try to find suitable treatments which promote gene expression of TIMPs with the DNA sequence information of regulatory region of TIMP2b gene.
Using the BAC library of cloned Japanese flounder genomic DNA, we isolated two BAC clone containing TIMP2b gene. As a result of investigation of 5' flunking region of TIMP2b gene, we found a cis-element called CRE-BP1, which is known to response to stress such as UV, heat-shock, and osmotic pressure. To investigate the function of CRE-BP1, the luciferase expression vector which contains a series of deleted 5' flunking region of TIMP2b.
The 5' flunking region of TIMP2b responded to hyper-osmotic stress, but do not to hypo-osmotic, heat-shock, and UV stress. These results suggested that the possiblity to elongate the toughness of fish meat durng chilled storage.
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