The immunochemical assay of the carp zinc-binding protein

• Main Authors: Jou-Ping Hsu, 許柔平
• Other Authors: Sen-Shyong Jeng
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/72631260846762115436

碩士 === 國立臺灣海洋大學 === 食品科學系 === 97 === 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.