具特定交互作用之重組蛋白對之研究：設計、製備及鑑定

Abstract

依照高分子鏈的結構可將高分子分類為以下四類：直鏈狀高分子、支鏈狀高分子、交聯狀高分子、網狀高分子。其中交聯狀高分子與網狀高分子常透過化學鍵交聯的方式在高分子鏈的交聯位點的部分以共價鍵鍵結的方式連接高分子鏈。在本研究中，預計利用基因工程的方式設計出一對具有特殊交互作用力的重組蛋白對，並期待在未來可將此對蛋白對用以取代交聯狀高分子結構中的交聯位點部分，賦予高分子材料在生物醫材領域上的應用更多的彈性與發展性。 利用基因工程的方式，將所需目標胺基酸鏈之基因序列插入至目標載體pET32b (+)後，轉殖入大腸桿菌表達系統中進行大量的重組蛋白表達。目標胺基酸鏈之基因模板，參考自兩個位於免疫細胞上的穿膜蛋白—CD40與CD154的膜外胺基酸序列，後續將分別表達出ReCD40與ReCD154兩個重組蛋白供實驗研究之用。 根據本實驗研究的實驗結果顯示，所設計的ReCD40與ReCD154重組蛋白對，在體外的環境下，彼此間確實具有特殊的交互作用力可使彼此連結。在未來，將會建立在ReCD40與ReCD154之間的連接關係的基礎上，將兩個重組蛋白修飾至高分子鏈上，作進一步的新型生醫材料的開發。

According to the structure of polymer chains, polymers can be classified into the following four categories: linear polymers, branched polymers, cross-linked polymers, and network polymers. The cross-linked polymer and the network polymer link the polymer chains from one backbone to another via covalent bonds between the polymer molecules to form a connected of three-dimensional structure. In this study, we expect to express a pair of recombinant proteins with specific interaction by genetic engineering, and look forward to using this pair of proteins to develop a new polymer crosslink mechanical in the future. This idea would give polymer materials a more flexibility development in the application of biomedical materials field. We insert the gene sequence into our target vector pET32b (+) by gene cloning, and then transfer it into the E. coli expression system for recombinant protein expression. The gene template of our target amino acid chain is referenced to the extramembrane amino acid sequences of two transmembrane proteins, CD40 and CD154, located on immune cells. According to the results of our study, ReCD40 and ReCD154 recombinant protein pairs do have a specific interaction to connect each other in vitro. In the future, we will base on the connection between ReCD40 and ReCD154. And both of them would be modified onto the polymer chains for further development of new biomedical materials.