| 牛源化脓隐秘杆菌PLO蛋白的原核截短表达及其溶血活性检测 |
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| 引用本文： | 周玉龙,高小兵,范春玲,王密,于红欣,侯喜林,朴范泽,朱战波.牛源化脓隐秘杆菌PLO蛋白的原核截短表达及其溶血活性检测[J].中国预防兽医学报,2012,34(3):197-200. |
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| 作者姓名： | 周玉龙 高小兵 范春玲 王密 于红欣 侯喜林 朴范泽 朱战波 |
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| 作者单位： | 1. 黑龙江八一农垦大学动物科技学院,黑龙江大庆,163319
2. 大庆禾丰八一农大动物科技有限公司,黑龙江大庆,163316 |
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| 基金项目： | 黑龙江省科技厅项目(GA09B301-2);黑龙江省农垦总局项目(HNK11A-08-01-07) |
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| 摘 要： | 为研究牛源的化脓隐秘杆菌溶血素(PLO)生物学功能,本研究应用PCR方法扩增牛源化脓隐秘杆菌(A.pyogenes)PLO全基因序列,通过生物信息学方法分析其与猪源A.pyogenes的PLO蛋白差异,并构建了溶血功能区重组表达质粒pQE30-PLO585,在E.coli XL1Blue中用IPTG诱导表达。结果表明,扩增到PLO蛋白基因ORF为1 605 bp,编码535个氨基酸,与猪源PLO的核苷酸序列同源性为97.4%,氨基酸同源性为97.2%,在生物学活性功能区没有发生改变。表达的PLO蛋白溶血功能区重组蛋白能够被阳性血清识别,而且具有溶解绵羊红细胞的活性,产生β溶血现象。本研究获得了牛源A.pyogenes截短重组PLO蛋白,并证明PLO具有β溶血功能与较好的抗原性。
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| 关 键 词： | 牛化脓隐秘杆菌 PLO蛋白 生物学功能 |
Prokaryotic expression of truncated pyolysin protein and its hemolytic activity of Arcanobacterium pyogenes isolated from cattle |
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| ZHOU Yu-long
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GAO Xiao-bing
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FAN Chun-ling
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WANG Mi
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YU Hong-xin
,
HOU Xi-lin
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PIAO Fan-ze
,
ZHU Zhan-bo.Prokaryotic expression of truncated pyolysin protein and its hemolytic activity of Arcanobacterium pyogenes isolated from cattle[J].Chinese Journal of Preventive Veterinary Medicine,2012,34(3):197-200. |
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| Authors: | ZHOU Yu-long GAO Xiao-bing FAN Chun-ling WANG Mi YU Hong-xin HOU Xi-lin PIAO Fan-ze ZHU Zhan-bo |
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| Institution: | 1(1.Animal Technology College,Heilongjiang Bayi Agricultural University,Daqing 163319,China;2.Daqing Wellhope Bayi Nongda Technology Co.Ltd,Daqing 163316,China) |
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| Abstract: | To express and identify the hemolytic activity of pyolysin(PLO) of Arcanobacterium pyogenes isolated from cattle,the complete gene of PLO was amplified by PCR,and the PLO gene from cattle and swine A.pyogenes strains were analyzed by bioinformatics methods.The expression recombinant plasmid with truncated PLO585 was constructed,and expressed by IPTG induction in E.coli XL1Blue.The results shown that the complete ORF of PLO gene was 1 605 bp encoded 585 amino acids.The homology of the nucleotide and the amino acids sequences of the PLO were 97.4% and 97.2% between bovine and swine A.pyogenes strains,respectively,without change in biologic activity domains.The truncate PLO containing haemolysis domain was expressed in E.coli which was able to react with positive serum and cursed β-hemolysis of sheep erythrocyte. |
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| Keywords: | bovine Arcanobacterium pyogenes pyolysin protein biological function |
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