串联启动子调控下的t-PAcDNA乳腺定位表达

摘要

The specific expression plasmid pSVL-WAPβ-PA, c ontaining t-PA gene, was constructed by linking up bovine β-Cansein and WAP p romoter. The three plasmids, pSVL-βb-PA，pSVL-WAP-PA and pSVL-WAPβ- PA，were mixed with Liposome and injected directly into pregnant rabbit mammary gland duct. Results of fibrin clot lysis assay showed that the three plasmids co uld express t-PA in rabbit milk. The expression level day 390， 440，450 μg*L－1 respectively reached the highest on the 4th day. This has laid a foundation for further study on the regulation of specific expression in mammary gland and the establishment of t-PA mammary gla nd bioreactor.%由牛β-Casein启动子与大鼠WAP启动子串联构建了t- PAcDNA 乳腺定位表达载体pSVL-WAPβ-PA，将3种表达载体pSVL-βb-PA,pSVL-WAP-PA及pSV L-WAPβ-PA分别注入怀孕家兔乳腺导管中，溶圈试验结果显示，3种表达载体均能在家兔乳腺中表达，且以分娩后第4 d的乳汁中表达水平最高，分别为390，440，450 μg*L －1．本试验为进一步研究t-PA基因乳腺定位表达调控和建立t-PA乳腺生物反应器奠定了基础.