转人溶菌酶基因小鼠乳腺生物反应器的建立

目的 研究人溶菌酶（human lysozyme，hLYZ）动物乳腺生物反应器制备的可行性。方法 将hLYZ基因与动物乳腺特异表达载体p205C3连接，所得重组载体p205C3-hLYZ用显微注射法建立转基因小鼠。结果 共出生了136只F0代小鼠，PER和Southern杂交检测基因整合阳性率分别为5．15％（2♀5♂）和2．94％（1♀3♂）。Western印迹检测结果表明，分泌在小鼠乳汁中的表达产物与正常hLYZ具有相同的分子量。目前转基因小鼠已经繁殖到B代，每一代基因整合阳性母鼠的乳腺均表达hLYZ，乳汁中的表达量最高达750mg/L。斑点杂交试验证明，表达有较强的组织特异性，除在乳腺表达外，仅在脾脏和小肠有一定的异位表达。结论 成功建立了hLYZ小鼠乳腺生物反应器。

Generation of transgenic mice expressing human lysozyme in mammary gland

OBJECTIVE: To evaluate the feasibility of generating animal mammary gland bioreactors expressing human lysozyme (hLYZ). METHODS: The recombinant vector p205C3-hLYZ, as a result of connecting the hLYZ cDNA with the mammry gland expression vector p205C3, was used to generate transfer genic mice by microinjection. RESULTS: A total of 136 F0 mice were obtained, of which 7 (2 females and 5 males) and 4 (1 females and 3 males) were found to contain the transfer-gene by PCR and Southern blotting respectively. The results of Western blotting indicated that the expressed protein had the same molecular weight as that of normal hLYZ. From the F1 generation on, the mice mated only with their brothers or sisters and a colony of F7 transgenic mice was obtained. Among the offspring, the female transgenic mice maintained and expressed the transfer-gene stably with an expression level as high as 750 mg/L. The expressed protein had strong tissue specificity, and in addition to the mammary glands, some degree of ectropic expression in the spleens and intestines of the transgenic mice was confirmed by dot blotting assay. CONCLUSION: These data indicate that the mice mammary gland bioreactors expressing hLYZ have been successfully generated.