2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/37965The objectives of this proposal are to investigate the catalytic reaction of novel oxidoreductase (putative short chain dehydrogenase) :（NCBI Nos. CAD54662(Zebrafish) and NP_055049(human)）to characterize its physiological function in zebrafish embryos model system. In order to investigate its catalytic reaction, the specific aims include: 1. to obtain the entire cDNA by using PCR; 2. to isolate and purify enzymes by using E. coli expression system; and 3. to establish the kinetic studies system in vitro. In order to characterize its physiological function, the specific aims include: 1.to set-up the Northern analysis; 2.to set-up the Western analysis; and 3.to perform RNA interference or RNA overexpression in zebrafish. In general, by using the approaches mentioned above, the novel enzymes will be identify and characterize in both molecular and physiological level.本計劃之目的是確認新穎的氧化還原酵素(一種假設性的短鏈去氫., short chain dehydrogenase)，其NCBI 編號為CAD54662(Zebrafish)以及NP_055049(human)之催化特 性及生理功能。為了達成此一目的，本計劃之進行可分成二個主要目標，分別是第一、 分析該酵素催化的反應；第二、觀察此類酵素在生物體之分佈及其效應。 為了分析該酵素催化的反應，本計劃將包括以下之特定目標： 1. 利用PCR 得到全長之cDNA； 2. 利用表現質體表現並純化該酵素； 3. 利用酵素動力學分析該酵素之特性。 為了觀察此類酵素在生物體之分佈及其效應，本計劃將包括以下之特定目標： 1.利用北方墨點分析標定酵素在生物體之分佈; 2.利用西方墨點分析標定酵素在細胞之分佈; 3.利用斑馬魚的胚胎糸統進行活體內之功能測試。 透過以上之分析，將可對上述二個新穎酵素之催化特性及生理功能有進一步的認識。酵素動力學斑馬魚的胚胎糸統RNA interferenceZebrafishenzyme kineticsProject