[1]贡莎莎,孟庆玲,乔军,等.少孢节丛孢菌几丁质酶AO-801基因的分子特征分析及其多克隆抗体制备[J].江苏农业科学,2018,46(10):23-27.
　Gong Shasha,et al.Molecular characterization analysis and polyclonal antibody preparation of chitinase AO-801 gene in Arthrobotrys oligospora Xinjiang isolate[J].Jiangsu Agricultural Sciences,2018,46(10):23-27.
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少孢节丛孢菌几丁质酶AO-801基因的分子特征
分析及其多克隆抗体制备()

《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:
第46卷
期数:
2018年10期
页码:
23-27
栏目:
生物技术
出版日期:
2018-05-20

文章信息/Info

Title:
Molecular characterization analysis and polyclonal antibody preparation of chitinase AO-801 gene in Arthrobotrys oligospora Xinjiang isolate
作者:
贡莎莎1;  孟庆玲1;  乔军1;  钟文强1;  陈英1;  才学鹏2
1.石河子大学动物科技学院，新疆石河子 832003； 2.中国农业科学院兰州兽医研究所，甘肃兰州 730046
Author(s):
Gong Shasha; et al
关键词:
少孢节丛孢菌; 几丁质酶; 原核表达; 蛋白纯化; 多克隆抗体
Keywords:
-
分类号:
S432.4+5；Q785
DOI:
-
文献标志码:
A
摘要:
为了研究捕食线虫性真菌——少孢节丛孢菌几丁质酶的生物学功能，对少孢节丛孢菌XJ-A1几丁质酶AO-801基因进行克隆和分子特征分析，通过大肠杆菌表达AO-801重组几丁质酶，利用镍柱纯化技术纯化目的蛋白，免疫小鼠后制备AO-801几丁质酶多克隆抗体。结果显示，几丁质酶AO-801基因序列与少孢节丛孢菌标准株［Arthrobotrys oligospora Fres.，美国模式培养物集存库(American type culture collection，简称ATCC)24927］的核苷酸序列同源性为95.96%，氨基酸序列的同源性为97.34%。几丁质酶AO-801属于糖苷水解酶18家族，具有典型的几丁质酶催化区保守序列SIGGW和LDGVDVDWE，无信号肽序列，其二级结构以无规则卷曲、α螺旋和β折叠为主要结构元件，三级结构中有(α/β)8的圆桶形结构。十二烷基硫酸钠聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis，简称SDS-PAGE)和蛋白质印迹法(Western Blot)分析表明，重组几丁质酶蛋白的分子量约为59ku，与预期大小一致，与少孢节丛孢菌阳性血清能发生特异性反应。间接酶联免疫吸附测定(enzyme linked immunosorbent assay，简称ELISA)法的检测结果显示，获得的抗AO-801多克隆抗体血清效价达到 1 ∶ 25 600。
Abstract:
-

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期：2017-08-21
基金项目：国家自然科学基金(编号：31460654、31260601)；新疆维吾尔自治区研究生科研创新项目(编号：XJGRI2015038)。
作者简介：贡莎莎(1991—)，女，新疆博乐人，硕士研究生，主要从事动物寄生虫学研究。E-mail：1558560806@qq.com。
通信作者：孟庆玲，博士，教授，主要从事动物寄生虫学研究。E-mail：xjmqlqj@163.com。
更新日期/Last Update: 2018-05-20