利用生物資訊資料庫NCBI（National Center for Biotechnology Information）中的搜尋比對工具BLAST（Basic Local Alignment Search Tool），搜尋比對到一組相對應的斑馬魚（編號NM_131852/NP_571927）與人類（編號NM_003359/NP_003350）尿核雙磷酸葡萄糖去氫(簡稱UGDH)。此斑馬魚尿核雙磷酸葡萄糖去氫(簡稱zUGDH) cDNA之開放閱讀框架（Open Reading Frame）含有1482個鹼基對，可轉譯成493個胺基酸，估計分子量54.480kDa；而人類尿核雙磷酸葡萄糖去氫(簡稱hUGDH) cDNA之開放閱讀框架含有1485個鹼基對，可轉譯成494個胺基酸，估計分子量55.024kDa。藉由pQE表達質體系統，將hUGDH表達與純化出來，將可成為酵素分析之基準。此外，利用此放大的cDNA可作為探針，進行北方墨點分析(Northern Blotting)，發現此酵素在大鼠的腦、心、肝、胃、睪丸皆有表現。經過NCBI網站之UniGene搜尋大鼠UGDH在各組織的表現，發現在腦、肝、消化系統、生殖系統有表現，與我們實驗結果稍有不同。這些實驗結果可提供hUGDH結構功能之後續相關研究的基礎。
In this study, the search tool BLAST (Basic Local Alignment Search Tool), based on NCBI (National Center for Biotechnology Information) was used against nucleotide and protein database. The analysis found a UDP-glucose dehydrogenase (UGDH) of zebrafish (NCBI accession number NM_131852/ NP_571927) and human (NCBI accession number NM_131852/ NP_571927). The UGDH cDNA of zebrafish and human were successfully amplified with a continuous open reading frame of 1482 and 1485 bps encoding a protein of 493 and 494 amino acids with a calculated molecular mass of 54.480 and 55.024 kDa respectively. The human UDP-glucose dehydrogenase expression vector (pQE-9) with a 6-Histidine tag for specific purification was also constructed. Northern Blotting analysis showed that hUGDH is expressed in the rat tissues, such as brain, heart, liver, stomach and testis. These results provide the basis for the further studies of structure-function relationship of the human UDP-glucose dehydrogenase.
