基于噬菌体展示技术的赭曲霉毒素A高密度模拟表位的表达研究

食品科学 ›› 2012, Vol. 33 ›› Issue (19): 188-192.

基于噬菌体展示技术的赭曲霉毒素A高密度模拟表位的表达研究

陈兴龙，徐玲，刘仁荣*，裘雪梅，朱立鑫

江西科技师范学院生命科学学院

收稿日期:2011-08-28 修回日期:2012-08-13 出版日期:2012-10-15 发布日期:2012-09-17
通讯作者: 刘仁荣 E-mail:lilirenrong@hotmail.com
基金资助:
国家自然科学基金项目;江西省自然科学基金

High Density Expression of Ochratoxin A Mimotope Based on Phage Display Technology

Received:2011-08-28 Revised:2012-08-13 Online:2012-10-15 Published:2012-09-17

摘要/Abstract

摘要： 通过噬菌体展示技术展示赭曲霉毒素A(ochratoxin A，OTA)模拟表位。将带有OTA模拟表位序列的寡核苷酸双链克隆至载体pC89-COTA。利用噬菌体展示技术，通过优化辅助噬菌体感染复数、IPTG加入的时间与剂量等培养条件获得表达有高密度OTA模拟表位的噬菌体。再通过酶联免疫吸附检测(ELISA)方法比较不同的条件下的表达效果，探索最优表达条件。结果表明：成功获得高密度表达OTA模拟表位的噬菌体，辅助噬菌体感染复数与IPTG加入量对模拟表位表达量影响不大，IPTG加入时间对模拟表位表达量有较大影响，优化后模拟表位表达量大大高于优化前。

关键词: 噬菌体展示, 噬菌粒, 酶联免疫吸附检测(ELISA), 赭曲霉毒素A

Abstract: Ochratoxin A (OTA) mimotope was displayed on phage surface. Two complementary oligonucleotides which contained OTA mimotope DNA sequences were cloned into vector pC89S4. Helper phage MOI (multiplicity of infection) and IPTG addition were optimized to gain phages with high density expression of OTA mimotope through phage display technology. The expression efficiency was measured by ELISA analysis. The result showed that high density OTA mimotope phages were obtained successfully. Helper phage MOI and IPTG concentration showed little influence the expression of OTA mimotope. However, IPTG addition at different time points had great effects on the expression of OTA mimotope. The expression efficiency was greatly improved after optimization.

Key words: phage display, phagemid, ELISA, ochratoxin A

中图分类号:

TS207

陈兴龙，徐?玲，刘仁荣*，裘雪梅，朱立鑫 . 基于噬菌体展示技术的赭曲霉毒素A高密度模拟表位的表达研究[J]. 食品科学, 2012, 33(19): 188-192.

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