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当前位置: 首页 > 文章 > 抗黄曲霉毒素B_1单链抗体的表达载体的比较 食品科学 2010,31 (9) 171-176
Position: Home > Articles > Comparison of pET Vectors for the Expression of Single Chain Antibody for Aflatoxin B_1 FOOD SCIENCE 2010,31 (9) 171-176

抗黄曲霉毒素B_1单链抗体的表达载体的比较

作  者：
杨炼;刘自琴;刘蓉;陈海琴;陈卫;张灏
单  位：
江南大学食品学院食品科学与技术国家重点实验室
关键词：
黄曲霉毒素;单链抗体;表达;大肠杆菌;pET
摘  要：
目的:比较4种不同的pET载体在表达抗黄曲霉毒素B1(AFB1)的单链抗体(scFv)克隆的特点,确定用以表达scFv-H4的合适载体。方法:将目的基因H4分别克隆到载体pET20b、pET22b、pET28a和pET32a上,分别转入大肠杆菌BL21(DE3)或Origami(DE3)中,比较诱导过程中细胞的生长状况和诱导结束后细胞破碎液中scFv-H4的生物活性以及细胞各部分包涵体。结果:pET20b和pET22b能表达具有生物活性的scFv-H4,这些具有生物活性的蛋白主要存在于周质中;pET28a和pET32a不能表达有生物活性的scFv-H4,而pET32a仅能在细胞质中产生大量的包涵体。结论:pET22b可能是表达AFB1的单链抗体较优的表达载体。
译  名：
Comparison of pET Vectors for the Expression of Single Chain Antibody for Aflatoxin B_1
作  者：
YANG Lian,LIU Zi-qin,LIU Rong,CHEN Hai-qin,CHEN Wei,ZHANG Hao*(State Key Laboratory of Food Science and Technology,School of Food Science and Technology,Jiangnan University,Wuxi 214122,China)
关键词：
aflatoxin;single chain fragment;expression;Escherichia coli;pET
摘  要：
Objective:Recombinant antibody expression in microorganism greatly facilitated the affinity and stability studies of recombinant antibodies,but previous studies had less reports about the selection of vectors.A suitable expression vector is important during the expression of the single chain antibody(scFv-H4) for aflatoxin B1.Methods:Four pET vectors such as pET20b,pET22b,pET28a and pET32a were employed in the functional expression of scFv-H4.Results:pET20b and pET22b vectors were able to express the functional antibody in periplasm.The vector of pET28a was not suitable for the expression of scFv-H4 due to lack of detectable scFv-H4 in total protein.Moreover,the detectable scFv-H4 was only expressed in inclusion body through pET32a expression system.Conclusion:pET22b was an excellent vector for the expression of scFv-H4,which might be a good reference for similar protein expression and be useful in the later study of scFv-H4 affinity and stability.

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