Wu Xiaojie, Lin Yanli, Xi Yongyi, Shao Zhenlu, Zhou Yanrong, Liu Fang, Chen Hongxing
Cell Engineering Department, Beijing Institute of Biotechnology, No. 20 Dongdajie Street, Beijing, 100071, People's Republic of China.
Biotechnol Lett. 2014 Jun;36(6):1197-202. doi: 10.1007/s10529-014-1476-7. Epub 2014 Feb 22.
Human lysozyme (hLYZ) has important potential applications as antimicrobial medicine and food additive. To develop a robust expression vector that ensures expression of large amounts of hLYZ in milk, here a 26,267 bp chimeric mouse whey acidic protein (mWAP)::hLYZ cassette was constructed and used as a mammary gland-specific expression vector, in which a 3,010 bp genomic sequence in the 24,466 bp mWAP gene locus was substituted by a 4,811 bp genomic sequence of hLYZ, exactly from the start codon to the stop codon. Corresponding transgenic mice were generated, and enzymatically-active hLYZ was expressed at 18.4-35 g l(-1) in the milk of most transgenic mouse lines. Our transgenic mice carrying chimeric mWAP::hLYZ represent a model system for cost-effective production of hLYZ.
人溶菌酶(hLYZ)作为抗菌药物和食品添加剂具有重要的潜在应用价值。为了构建一种强大的表达载体,以确保在乳汁中大量表达hLYZ,本文构建了一个26267 bp的嵌合小鼠乳清酸性蛋白(mWAP)::hLYZ盒,并将其用作乳腺特异性表达载体,其中24466 bp的mWAP基因座中的3010 bp基因组序列被hLYZ的4811 bp基因组序列取代,该序列从起始密码子到终止密码子。生成了相应的转基因小鼠,大多数转基因小鼠品系的乳汁中均可表达酶活性hLYZ,表达量为18.4 - 35 g l(-1)。我们携带嵌合mWAP::hLYZ的转基因小鼠代表了一种经济高效生产hLYZ的模型系统。