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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 expression characteristics of self-cloned 1.5kb hLYZ double-stranded cDNA. Methods The cDNA was subcloned into the eukaryotic expression vector pcDNA3 and mammary gland specific expression vector p205C3. The recombinant vector pcDNA3LYZ was transfected into COS-1 cells. The expression of hLYZ cDNA in transfected cells was detected by indirect immunofluorescence. The recombinant plasmid p205C3LYZ Mice were used to detect the expression of hLYZ cDNA in mice by Micrococcal lysis assay and dot hybridization assay. Results hLYZ cDNA was correctly expressed in transfected COS-1 cells. The expression of hLYZ in mice was 87μg / ml, and the expression of the target gene was well tissue-specific. Conclusion hLYZ cDNA can be efficiently expressed in COS-1 cells. The gene p205C3LYZ can be used in animal mammary gland bioreactor.