利用新型α-半乳糖苷酶进行B→O血型改造  

B to O blood group conversion by new type α-galactosidase

作　　者：王玲燕[1] 高红伟[1] 金泗虎[1] 李素波[1] 虞立霞[1] 鲍国强[1] 宫锋[1] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850

出　　处：《中国输血杂志》2011年第2期122-125,共4页Chinese Journal of Blood Transfusion

摘　　要：目的利用原核生物来源的α-半乳糖苷酶进行B→O血型改造研究。方法应用PCR的方法从脆弱类杆菌(Bacteriodes fragilis)中克隆了新型α-半乳糖苷酶基因,构建原核生物表达载体,转化大肠杆菌BL21(DE3),IPTG诱导α-半乳糖苷酶的胞内可溶性表达,应用重组酶的上清液进行B型红细胞(RBC)初步酶解试验。结果重组酶分子量约为64.6 kD,超声破碎的菌液上清中的酶活力约为2 U/ml。在26℃及pH 6.8的等渗柠檬酸-磷酸氢二钠缓冲体系中2,h内完全酶解200μl B型RBC需要30μl的上清液,酶解后的B型RBC与抗-B和抗-A单克隆抗体不凝集,流式细胞术检测结果显示酶解后B型RBC的血型抗原呈现O型特征。结论重组的α-半乳糖苷酶可以有效的将B型RBC转变为O型RBC。Objective To study the B to O blood group conversion by α-galactosidase from procaryote.Methods The α-galactosidase gene was cloned from Bacteriodes fragilis by PCR and inserted into the procaryotic expression vector.We induced soluble α-galactosidase by IPTG and used the ultrasonic disruption supernatant of E.coli to digest the B group red blood cells.Results The activity of α-galactosidase was about 2 U/ml and the molecular weight of α-galactosidase was about 64.6 kD.B antigen was cleaned within 2 hours by 30 μl supernatant at 26℃,pH 6.8 completely.The enzymatically converted red blood cells did not react with anti-B and anti-A monoclonal antibodies.Conclusion Group B red blood cells could be effectively converted to group O by recombinant procaryote α-galactosidase.

关 键 词：Α-半乳糖苷酶 血型改造 B抗原 

分 类 号：R457.11[医药卫生—治疗学]