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鸡输卵管表达的重组人溶菌酶的鉴定
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朱秋菊1 , 孙怀昌1 , 赵敏苏1, 王永娟1, 李国才2
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（1. 扬州大学 兽医学院, 江苏 扬州225009； 2. 扬州大学 医学院, 江苏 扬州225001）
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摘要：
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用直接结晶法从重组载体注射鸡蛋清中提取重组人溶菌酶（rhLYZ），并对其理化及生物学特性进行了鉴定。结果显示，rhLYZ能被hLYZ特异抗体识别；纯化的rhLYZ和天然hLYZ的热稳定性无显著差异，二者在pH3.0～11.0范围内均具有活性， pH为7.0时活性最强。采用最小抑菌浓度法测定了含rhLYZ蛋清和纯化rhLYZ对12种常见细菌的溶菌活性，二者对除有鞭毛伤寒杆菌和鳖嗜水气单胞菌以外的参试菌均有不同程度的抑制作用。表明，表达在重组载体注射鸡蛋清中的rhLYZ具有与天然hLYZ十分相似的分子质量、热稳定性、pH活性范围和溶菌谱。 |
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关键词：
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重组人溶菌酶； 特性； 鉴定 |
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中图分类号：S 852．42 文献标识码：
A 文章编号：1673-4696(2006)03-0234-05 |
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Identification of recombinant human lysozyme expressed in egg white |
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ZHU Qiu-ju1, SUN
Huai-chang1, ZHAO Min-su1, WANG
Yong-juan1, LI Guo-cai2 |
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(1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
2. College of Medicine, Yangzhou University, Yangzhou 225001,China) |
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Abstract: To identify the recombinant human lysozyme expressed in egg white, the lysozyme was purified from egg white of laying hens before and after injection with pOV2LYZ vector.
Western-blotting analysis of the purified product showed that rhLYZ was able to be recognized by
hLYZ-specific antibody with a molecular weight identical to that of the natural hLYZ in human colostrums. Colorimetric assay using
RBB-R-labelled Micrococcus luteus as the substrate showed that the rhLYZ had a similar
thermo-stability, and was active at pH3.0-11.0 with the highest activity at pH7.0. The tube test for minimum inhibition concentrations showed that the rhLYZ had a variable bactericidal effect on 10 different bacteria. The results suggested that the rhLYZ expressed in egg white of laying hens injected with the vector had similar biochemical properties to that of the natural hLYZ. |
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Key words: recombinant human lysozyme; characteristic; identification |
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