摘要
为了研制动物乳腺生物反应器,用中国奶山羊β-乳球蛋白基因的5’-、3’-侧翼区和β-酪蛋白基因的第1内含子构建动物乳腺特异表达载体p205C3。细胞表达试验结果显示,p205C3载体能够指导lacZ基因在SV40 T基因转化的山羊乳腺上皮细胞中表达,不能指导该基因在COS-1细胞和山羊成纤维细胞中表达。将人激肽释放酶(hKLK1)cDNA克隆入p205C3载体,用获得的重组载体p205C3KLK1注射哺乳期小鼠,然后取其心、肝、脾、肺、肾、脑、肌肉、胰腺和乳汁,酶活性测定结果显示,乳汁中的酶活性高达255.65 U·mL-1,其他组织中的酶活性与正常小鼠无差异。结果表明:p205C3载体不仅能有效地指导外源基因在山羊乳腺细胞和小鼠乳腺中表达,而且表达具有较好的组织特异性,可以用于动物乳腺生物反应器的研制。
To generate mammary gland bioreactors, the 5'-and 3'-flanking regions of β-lactoglobulin (BLG) gene and the first intron of β-casein gene of Chinese dairy goat were used to construct a mammary gland-specific expression vector p205C3. The vitro expression experiments showed that the vector was able to direct expression of lacZ gene in SV40 T-transformed goat mammary gland epithelial cells, but not in goat fibroblast and COS-1 cells. Human kallikrein (hKLKl) cDNA was subcloned into the p205C3 vector and the recombinant vector was injected into lactational mice. The KLK1 activities up to 255 U·mL-1were detected in the milk samples, but not in the heart, liver, spleen, lung, kidney, brain, pancreas and muscle of the mice. These results indicate that the p205C3 vector can be used as an expression vector for development of mammary gland bioreactors.
出处
《江苏农学院学报》
CSCD
2003年第4期1-4,共4页
Jiangsu Agricultural Research
基金
国家教育部骨干教师培养计划项目(2000-06)
