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The immunochemical assay of the carp zinc-binding protein
|Authors: ||Jou-Ping Hsu|
|Contributors: ||NTOU:Department of Food Science|
common carp;carp zinc-binding protein;immunochemical assay
|Issue Date: ||2011-06-30T08:12:15Z
|Abstract: ||摘要 鯉魚消化道組織常具有高濃度之鋅，未見於其他水產生物。鯉魚消化道組織之高鋅主要來自於一種位於結締組織細胞plasma membrane上之「鋅結合蛋白質」，其分子量為43kDa。此「鯉魚鋅結合蛋白質」已製成多株抗體。為了解鯉魚與其他魚類各種組織中 「鯉魚鋅結合蛋白質」之存在與含量，因此以此抗體發展免疫化學分析方法，進行檢測不同生物組織之「鯉魚鋅結合 蛋白質」。本研究所發展之免疫化學分析方法，可特異性地檢測「鯉魚鋅結合蛋白質」之量，其範圍為2-40 ng。應用上述免疫化學方法之分析，可以 得知，鯉魚之消化道組織、腎、頭腎、脾與血液中有較高「鯉魚鋅結合 蛋白質」含量，分別為2、1、12、7及1 mg/(g fresh tissue)；而 肝胰臟及肌肉中，則含量較低 (<0.13 mg/ g fresh tissue )。目前已檢測之魚類，包含鯽魚、草魚、鰱魚和吳郭魚等。鯽魚其腎、頭腎和脾中有較高「鯉魚鋅結合蛋白質」含量 (分別為0.15、0.21及0.66 mg/ [ g fresh tissue ])。草魚、鰱魚和吳郭魚其各組織中，幾乎沒有「鯉魚鋅 結合蛋白質」存在 (<0.04 mg/ g fresh tissue )。經由餵食「一般飼料」及「高鋅飼料」鯉魚，各組織之鋅及「鯉魚鋅結合蛋白質」之比較，得知餵食「高鋅飼料」鯉魚之消化道組織和頭腎不但鋅濃度增加，組織中的「鯉魚鋅結合蛋白質」也增加。然而，血液中之鋅雖無顯著變化，但其「鯉魚鋅結合蛋白質」含量卻明顯增加。利用報告所發展之「鯉魚鋅結合蛋白質」免疫化學分析法，應可廣泛了解此蛋白質在不同生物之存在及分布。另外，以此免疫化學方法測定鯉魚組織中「鯉魚鋅結合蛋白質」在各種不同環境或條件下如何變化，對於了解「鯉魚鋅結合蛋白質」的功能以及高鋅在鯉魚之生理意義，可能極為有用。|
Abstract High concentration of zinc was found in the digestive tract tissue of common carp which was not seen in other fish species. A zinc-binding protein with a molecular weight of 43kDa located on the plasma membrane of the connective tissue cells was responsible for the high zinc in common carp. Using the “carp zinc-binding protein” as antigen, polyclonal antibody of the protein was prepared. An immunochemical assay method using the antibody against the “carp zinc-binding protein” to quantitatively determine the amount of the protein in fish tissue was developed to study the existence of the protein in fish tissue. The immunochemical assay method developed could specifically detect a amount of “carp zinc-binding protein” between 2-40 ng. Based on this immunochemical assay, it was found that common carp had high amount of “carp zinc-binding protein” in its digestive tract tissue, kidney, head kidney, and blood with values of about 2, 1, 12, 7, and 1 mg/(g fresh tissue), respectively; but lower amount in its heaptopancreas and muscles (<0.20mg/ g fresh tissue ). In the other fishes, crucian carp had high amount of “carp zinc-binding protein” in its kidney, head kidney and spleen, but grass carp, silver carp and tilapia have little or no “carp zinc-binding protein” (<0.04 mg/ g fresh tissue ) in their tissues. The zinc and “carp zinc-binding protein” in the tissues of common carp fed basal diet was compared with those fed high zinc diet. It was found that the amount of zinc and “carp zinc-binding protein” in the digestive tract tissue and head kidney increased in the common carp fed high zinc diet. The zinc concentration in the blood of the common carp didn’t increase after feeding high zinc diet, but the amount of “carp zinc-binding protein” increased. This immunochemical assay method might be used to study the existence and distribution of “carp zinc-binding protein” in other organism. Besides, using immunochemical assay to determine the change of the “carp zinc-binding protein” in common carp under different circumstances or conditions might be very useful in understanding the function of “carp zinc-binding protein” and its physiological meaning.
|Appears in Collections:||[食品科學系] 博碩士論文|
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