鸡输卵管表达的重组人溶菌酶的鉴定  

Identification of recombinant human lysozyme expressed in egg white

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作  者:朱秋菊[1] 孙怀昌[1] 赵敏苏[1] 王永娟[1] 李国才[2] 

机构地区:[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

出  处:《中国兽医科学》2006年第3期234-238,共5页Chinese Veterinary Science

基  金:江苏省高新技术发展计划项目(BJ2001315)

摘  要: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茵均有不同程度的抑制作用。表明,表达在重组栽体注射鸡蛋清中的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。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 analysts of the purified product showed that rhI.YZ 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 Zuteus as the substrate showed that the rhLYZ had a similar thermostabiIity, and was active at pH 3.0-11.0 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.

关 键 词:重组人溶菌酶 特性 鉴定 

分 类 号:S852.42[农业科学—基础兽医学]

 

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