人溶菌酶cDNA在COS-1细胞及小鼠乳腺的暂态表达

Temporary Expression of Human Lysozyme cDNA in COS-1 Cells and Murine Mammary Gland

目的研究自行克隆的1.5kb hLYZ双链cDNA的表达特性。方法将此cDNA亚克隆入真核表达载体pcDNA3及乳腺特异表达载体p205C3,重组载体pcDNA3LYZ转染COS-1细胞,用间接免疫荧光法检测转染细胞中hLYZcDNA的表达;重组载体p205C3LYZ注射哺乳期小鼠,用微球菌溶解试验和斑点杂交试验检测hLYZ cDNA在小鼠体内的表达。结果hLYZ cDNA在转染的COS-1细胞中得到了正确表达;小鼠体内表达并分泌到乳汁中的hLYZ达87μg/ml,且目的基因的表达具有较好的组织特异性。结论hLYZ cDNA可在COS-1细胞中有效表达,基因构件p205C3LYZ可以用于动物乳腺生物反应器研制。

Objective To study the characteristics of expression of self-cloned 1.5 kb hLYZ cDNA. Methods Subelone the 1.5 kb hLYZ cDNA into an eukaryotie expression vector pcDNA3 and mammary gland expression vector p205C3.Transfect COS-1 ceils with recombinant plasmid pcDNA3LYZ and detect the expression of hLYZ cDNA by IFA. Inject the mice during lactating period with recombinant plasmid p205C3LYZ and detect the expression of hLYZ cDNA in vivo of mice by mierococcal lysis assay and dot blot. Results hLYZ cDNA was correedy expressed in the transfected COS-1 celia. The mean expression level of hLYZ in milk of mice was 87 μg/ml. The expression of target protein was tissue-specific. Conclusion The self-cloned hLYZ cDNA may be used for the preparation and development of recombinant hLYZ. The recombinant plasmid p205C3LYZ may be used for the preparation of animal mammary gland bioreaetors.