灭活铜绿假单胞菌适体的筛选

被引：6

作者：

曾燕丽

兰小鹏

江丽

刘丰伟

李卫滨

机构：

[1] 福州总医院全军检验医学研究所

来源：

中国生物化学与分子生物学报 | 2009年 / 25卷 / 01期

关键词：

指数富集配基的系统进化技术; 铜绿假单胞菌; 适体;

D O I：

10.13865/j.cnki.cjbmb.2009.01.012

中图分类号：

R378 [病原细菌];

学科分类号：

摘要：

利用指数富集配基的系统进化(SELEX)技术,以灭活的铜绿假单胞菌为靶标,从体外合成的96 nt随机ssDNA文库中筛选与铜绿假单胞菌特异结合的适体.在第12轮和第14轮与其它假单胞菌属进行反筛策略,并进行了适体的结合亲和力的测定,再分别利用ClustalX、Mega2和Mfold sever软件分析适体的一级和二级结构.研究结果表明,经过15轮筛选,随机ssDNA文库与铜绿假单胞菌结合的A值从0.022上升到0.448.反筛与未反筛的结合A比值最高比为53倍.经过第15轮反筛后的24个阳性克隆子测序,根据软件分析,其可分成10个家族.每个家族都有其共同的保守序列(除第10个家族),其中有2条序列几乎完全一致(F23和F47),同源性达到97%,A值分别高达1.598和1.508,Kd为14.55和77.46 nmol/L,间接说明适体与铜绿假单胞菌的结合力明显增高.

引用

页码：90 / 97

页数：8

共 18 条

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