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为进一步探讨乙酰辅酶Ａ：胆固醇酰基转移酶的结构和功能的关系，经聚合酶链反应扩增得到的乙酰辅酶Ａ∶胆固醇酰基转移酶（ＡＣＡＴ）的Ｃ末端膜外结构域（氨基酸４８０～５４５）用Ｅｃｏｒｌ酶解，插入到表达载体ｐＧＥＸ２ＴＫ中，得到重组表达质粒ｐＧＥＸ２ＴＫ／ＡＣＡＴ（氨基酸４８０～５４５）。阳性重组子在大肠杆菌中经异丙基βＤ硫代半乳糖苷诱导表达谷胱甘肽转移酶－乙酰辅酶Ａ∶胆固醇酰基转移酶（氨基酸４８０～５４５），重组表达菌裂解上清液经谷胱甘肽ＳｅｐｈａｒｏｓｅＣＬ４Ｂ亲和柱纯化。十二烷基硫酸钠－聚丙烯酰胺凝胶电泳和Ｗｅｓｔｅｒｎｂｌｏｔｉｎｇ分析显示得到了较纯的谷胱甘肽转移酶－乙酰辅酶Ａ∶胆固醇酰基转移酶（氨基酸４８０～５４５）融合蛋白。乙酰辅酶Ａ∶胆固醇酰基转移酶的Ｃ末端膜外结构域（氨基酸４８０～５４５）的分离纯化为抗谷胱甘肽转移酶－乙酰辅酶Ａ∶胆固醇酰基转移酶（Ｃ未端片段）抗体的制备及其乙酰辅酶Ａ∶胆固醇酰基转移酶结构和功能关系的进一步研究打下了基础。
To further explore the relationship between the structure and function of acetyl-CoA: cholesterol acyltransferase, the C-terminal extracellular domain of acetyl-CoA: cholesterol acyltransferase (ACAT) amplified by polymerase chain reaction (amino acids 480-545 ) With Ecorl digestion, inserted into the expression vector pGEX 2TK, recombinant expression plasmid pGEX 2TK / ACAT (amino acids 480 ~ 545). Positive recombinant in Escherichia coli induced by isopropyl β D thiogalactoside glutathione transferase acetyl CoA: cholesterol acyltransferase (amino acids 480-545), the recombinant expression of bacterial lysate supernatant The liquid was purified by glutathione-Sepharose CL-4B affinity column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Westernbloting analysis showed that the more pure glutathione transferase-acetyl-CoA: cholesterol acyltransferase (amino acids 480-545) fusion protein was obtained. Isolation and Purification of the C-terminal Ectodomain (Amino Acids 480-545) of Acetyl-Coenzyme A: Cholesteryl Acyltransferase Preparation of Antibody Against Glutathione Transferase-Acetyl-CoA: Cholesteryl Acyltransferase (C-Terminal Fragment) And its acetyl-CoA: cholesterol acyltransferase structure and function of the further study laid the foundation.